WO2017146184A1

WO2017146184A1 - Laser endoscope device

Info

Publication number: WO2017146184A1
Application number: PCT/JP2017/006962
Authority: WO
Inventors: 明溝口; 光司田中; 裕二問山; 匡介田中; 淑杰王; 愛加垣内; 煌植崔; 一志木村
Original assignee: 国立大学法人三重大学
Priority date: 2016-02-23
Filing date: 2017-02-23
Publication date: 2017-08-31
Also published as: US11140318B2; CN108697315A; CN108697315B; JPWO2017146184A1; US20210227133A1; EP3420885A4; EP3420885A1; EP3420885B1; JP6738100B2

Abstract

The present invention is provided with: an imaging unit (10) that has an imaging head (11) to be inserted into the digestive tract (112) and images a living body by applying a laser to the digestive tract (112) via the imaging head (11); a control unit (50) for controlling the imaging head (11) to move inside the digestive tract (112); and an image processing unit (70) for processing an image captured by the imaging unit (10). The imaging unit (10) captures a plurality of imaging regions (P) to be imaged along with the movement of the imaging head (11) such that a portion of adjacent imaging regions (P1, P2) overlap, and the image processing unit (70) overlaps regions (Pa) in which the plurality of imaging regions (P1, P2) are overlapped to generate a composite image.

Description

Laser endoscope device

The present invention relates to a laser endoscope apparatus that images the inside of a living body.

Recently, as a method for confirming a lesion inside a living body (for example, the digestive tract), a method for confirming the presence or absence of a lesion such as a cancer cell by inserting an endoscope inside the living body is known.

As an example, Patent Document 1 describes a method of imaging a cell form inside a living body by staining a predetermined cell group inside the living body and then applying a multiphoton laser to the stained cell group. According to this method, since the stained cell group emits fluorescence by applying a multiphoton laser, a clear image of the cell morphology inside the living body can be obtained. Thereby, the presence or absence of lesions such as cancer cells can be accurately confirmed.

International Publication No. 2014/157703

However, in the method described in Patent Document 1, since the obtained image is an image of a local area inside the living body, the presence or absence of a lesion can be confirmed only within the imaged area. In addition, from the viewpoint of the person receiving the examination, it is impossible to grasp the presence or absence of a lesion outside the imaged region, so that anxiety remains.

The present invention solves the above-described problems, and an object of the present invention is to provide a laser endoscope apparatus capable of imaging a cell form inside a living body over a wide range without omission.

In order to achieve the above object, a laser endoscope apparatus according to an aspect of the present invention includes an imaging head that is inserted into a living body, and applies the laser to the living body via the imaging head. An imaging unit that images a living body, a control unit that controls the imaging head to move inside the living body, and an image processing unit that processes an image captured by the imaging unit, The plurality of imaging areas to be imaged in accordance with the movement of the imaging head are imaged such that adjacent imaging areas partially overlap, and the image processing unit superimposes the overlapping areas of the plurality of imaging areas. Generate a composite image.

According to this aspect, the cell morphology inside the living body can be imaged over a wide range without leakage.

For example, the imaging unit images the imaging region at a predetermined depth in a depth of 10 μm or more and 1000 μm or less from the mucosal surface inside the living body, and the image processing unit is configured to capture the composite image at the predetermined depth. May be generated.

According to this aspect, it is possible to image the cell morphology inside the living body at a predetermined depth of 10 μm or more and 1000 μm or less from the mucosal surface over a wide range without leakage.

For example, the control unit may control the movement of the imaging head so that the imaging head scans with a certain distance from the living body.

According to this aspect, the quality of a plurality of images obtained by imaging is stable, and a composite image with little unevenness can be obtained when a plurality of images are combined.

For example, the imaging head includes an objective lens arranged to face the living body, and a spacer provided around a space between the objective lens and the living body, and the control unit includes the spacer The fixed distance may be maintained by controlling the movement of the imaging head so as to contact the living body.

According to this aspect, the distance between the living body and the objective lens is constant, and the lens can be focused with high accuracy, so that a clear image can be obtained.

For example, the living body is a digestive tract, and the control unit controls the imaging head to move along the inner periphery of the digestive tract, and the imaging unit captures an image as the imaging head moves. A plurality of imaging regions that are overlapped with each other in the circumferential direction, and the image processing unit generates a panoramic image by superimposing the overlapping regions of the plurality of imaging regions on each other Also good.

According to this aspect, it is possible to comprehensively grasp the state of the inner wall of the digestive tract from the panoramic image.

For example, the living body is a digestive tract, the control unit controls the imaging head to rotate about the axis of the digestive tract, and the imaging unit captures an image as the imaging head rotates. Even if a plurality of imaging areas are imaged so that a part of the imaging areas adjacent to each other in the rotation direction overlap, the image processing unit may generate a panoramic image by superimposing the overlapping areas of the plurality of imaging areas. Good.

For example, the control unit may control the imaging head to revolve around the axis of the digestive tract.

According to this aspect, it is possible to capture images without omission while associating the imaging region with the position of the inner wall of the digestive tract.

For example, the control unit may control the imaging head to move in a spiral direction around the axis of the digestive tract.

According to this aspect, the inner wall of the digestive tract can be continuously imaged in a short time.

For example, the control unit controls the imaging head to move along the duct direction of the gastrointestinal tract, and the imaging unit captures a plurality of imaging areas to be imaged as the imaging head moves. Images may be picked up so that a part of the image pickup areas adjacent to each other in the duct direction overlap, and the image processing unit may generate the panoramic image by superimposing the overlapped areas of the plurality of image pickup areas.

According to this aspect, it is possible to grasp the position (coordinates) where a lesion exists in the duct direction of the digestive tract.

For example, the imaging head includes an objective lens, and a focus variable unit that can change a focal position of the objective lens in a depth direction from a cell surface of the living body, and the control unit includes the focus variable unit. By operating, the focal position is changed, the imaging unit images a plurality of imaging regions having different depths according to the change of the focal position, and the image processing unit is obtained by imaging of the imaging unit. By arranging a plurality of images corresponding to the focal position, a stereoscopic image inside the living body is obtained.

According to this aspect, it is possible to grasp the cell morphology inside the living body at a predetermined depth.

For example, the control unit changes the focus position at a first focus variable mode that changes the focus position at a first pitch, and at a second pitch that is smaller than the first pitch. And after the imaging in the first variable focus mode, there is a suspicious lesion in the image obtained by the imaging. The imaging may be performed in the second variable focus mode in the vicinity of the focal position when an image of a certain part is captured.

According to this aspect, comprehensive imaging can be performed while shortening the imaging time.

For example, the control unit stores in advance an image of normal cells in the absence of a lesion, and the image obtained in the first variable focus mode and the image of normal cells are at least of shape and brightness. One may be compared to determine the suspected lesion.

According to this aspect, the suspicion of the lesion can be objectively determined in a short time.

For example, when there is a diseased cell in the image obtained by the imaging unit, the control unit increases the output of the laser than during imaging, and increases the output of the laser to the diseased cell. The lesioned cells may be removed.

According to this aspect, the diseased cells can be removed early and reliably.

For example, the laser may be a multiphoton laser.

According to this aspect, tissue cells having a depth of about 1 mm from the surface inside the living body can be reliably imaged.

For example, the imaging device further includes a staining agent supply unit that supplies a staining agent for selectively staining cells in the living body to a chromatic color into the living body, and the imaging unit includes the staining agent supply unit. The cell group dyed by may be imaged.

According to this embodiment, a clear image of the stained cell group can be obtained.

For example, it further comprises a staining agent supply unit that supplies a staining agent for staining a cell group inside the living body into two or more chromatic colors that are different depending on the cell type, and the inside of the living body, The imaging unit may image the cell group stained with two or more colors by the staining agent supply unit.

According to this aspect, it is possible to obtain a clear image of a cell group stained with two or more colors. Further, for example, a plurality of tissues on the inner wall of the digestive tract can be simultaneously confirmed in one image.

Further, the laser endoscope apparatus according to one aspect of the present invention provides a staining agent for staining a cell group in a living body into two or more chromatic colors that are different depending on the type of the cell. And a imaging unit that images the cell group by applying a laser to the cell group stained by the staining agent supply unit.

According to this aspect, it is possible to obtain a clear image of a cell group stained with two or more colors. Further, for example, a plurality of tissue cells on the inner wall of the digestive tract can be simultaneously confirmed in one image.

For example, the staining agent may include two staining agents including both a curcumin class and acid red, or a staining reagent including curcumin class and an acid red color staining agent. It may be a staining agent.

According to this aspect, the cell group inside the living body can be surely stained in two colors, and a clear image can be obtained.

For example, the staining agent is based on a staining agent containing both curcumin (Curcumin) and Fast Green FCF (FastGreen FCF), or a staining agent containing Curcumin (Curcumin) and a staining agent containing Fast Green FCF (FastGreen FCF). Two dyes may be used.

In addition, the laser endoscope apparatus according to one aspect of the present invention specifically has a cancer cell peripheral cell group other than the cancer cells located around the cancer cell among the cell groups inside the living body. A staining agent supply unit for supplying a staining agent for coloring in the living body, and a cell around the cancer cell by applying a laser to a cell group inside the living body stained by the staining agent supply unit An image pickup unit for picking up an image capable of visually distinguishing the group.

According to this aspect, it is possible to obtain a clear image of cancer cell peripheral cell groups located around the cancer cell.

For example, the staining agent may be a staining agent including Rose Bengal.

According to this aspect, it is possible to reliably obtain a clear image of the cancer cell peripheral cell group located around the cancer cell.

A laser endoscope apparatus according to an aspect of the present invention includes an imaging head that is inserted into a living body, and an imaging unit that images the living body by applying a laser to the living body via the imaging head; A control unit for controlling the operation of the imaging head, the imaging head having an objective lens and a focus variable unit capable of changing a focal position of the objective lens in a depth direction from a cell surface of the living body. Then, the control unit changes the focal position by operating the focal point changing unit, and the imaging unit captures a plurality of images having different depths from the mucosal surface inside the living body according to the change of the focal position. Image the area.

For example, the imaging unit images the imaging region in a predetermined range of depths from 0 μm to 1000 μm from the mucosal surface inside the living body, and associates the captured image with the depth information. An image processing unit for storing and processing an image captured by the imaging unit, wherein the image processing unit arranges a plurality of images obtained by imaging of the imaging unit corresponding to the focal position; A stereoscopic image inside the living body may be generated.

According to this aspect, it is possible to grasp the cell morphology inside the living body at a depth of 0 μm or more and 1000 μm or less from the mucosal surface.

Furthermore, a staining agent supply unit that supplies a staining agent for selectively staining chromatic colors of cells inside the living body to the inside of the living body, and the imaging unit is stained by the staining agent supply unit. The obtained cell group may be imaged.

The imaging unit further includes a staining agent supply unit configured to supply a staining agent for staining a cell group inside the living body into two or more chromatic colors that are different depending on cell types, into the living body. May image the cell group stained with two or more colors by the staining agent supply unit.

Further, a stain for specifically staining a chromatic color of a cancer cell peripheral cell group other than the cancer cell located in the periphery of the cancer cell among the internal cell group of the living body, And the imaging unit may image the cancer cell peripheral cell group stained by the staining agent supply unit.

For example, the image processing unit generates a cross-sectional image of the stained cell group by cutting the plurality of images at a position including the stained cell group, and the control unit A suspected lesion may be determined based on the depth at which the cell group represented in the cross-sectional image is stained.

According to this aspect, the suspicion of a lesion can be objectively determined.

According to the present invention, it is possible to provide a laser endoscope apparatus capable of imaging a cell form inside a living body over a wide range without leakage.

In addition, according to the main configuration of the present invention, a minute lesion such as a very early cancer (diameter 0.2 mm to 1 mm) which is too small to be noticed by the current endoscope is not accidentally detected. It becomes possible to detect exhaustively over a wide range.

FIG. 1 is a schematic diagram showing the arrangement of colon cells, which is an example of the digestive tract. FIG. 2 is a diagram schematically showing cancer cells that develop in the digestive tract. FIG. 3 is a schematic view showing a state in which the inner wall of the digestive tract is imaged using a multiphoton laser microscope. FIG. 4 shows an image when epithelial cells and glandular cells are imaged using a multiphoton laser microscope after staining epithelial cells and glandular cells with a stain containing curcumin. FIG. 5 shows an image when capillaries and connective tissues are imaged using a multiphoton laser microscope after staining capillaries and connective tissues with a stain containing Acid Red. FIG. 6 shows an image when the inner wall of the digestive tract is double-stained with a stain containing curcumin and a stain containing acid red, and then the inner wall of the digestive tract is imaged using a multiphoton laser microscope. FIG. 7 shows an image when the inner wall of the digestive tract is imaged using a multiphoton laser microscope after staining the inner wall of the digestive tract with a stain containing rose bengal. FIG. 8 is a schematic diagram showing a state in which the inner wall of the digestive tract is imaged using a multiphoton laser microscope. Fig. 9 is a composite image of the inner wall of the digestive tract stained with a stain containing acid red FIG. 10A is a composite image of the inner wall of the digestive tract stained with a stain containing curcumin and a stain containing acid red. FIG. 10B is a diagram showing an example of a three-dimensional reconstruction of a panoramic image of the inner wall of the digestive tract FIG. 11 is a diagram showing a state after the insertion tube is inserted into the digestive tract in the laser endoscope apparatus according to the first embodiment. FIG. 12 is a diagram illustrating an example of a stain supply unit for supplying a stain in the laser endoscope apparatus according to Embodiment 1. FIG. 13A is a view showing a state in which the inner wall of the digestive tract is flattened using the laser endoscope apparatus according to Embodiment 1, and FIG. 13B is a view of the laser endoscope apparatus. Schematic diagram showing the end on the tip side FIG. 14 is a schematic diagram illustrating the entire endoscope in the laser endoscope apparatus according to the first embodiment. FIG. 15 is a block diagram showing a control configuration of the laser endoscope apparatus according to the first embodiment. FIG. 16 is a schematic diagram illustrating a state in which the inner wall of the digestive tract is imaged using the laser microscope according to the first embodiment. FIG. 17A is a diagram for explaining the operation of the laser endoscope apparatus according to the first embodiment. FIG. 17B is a diagram for explaining the operation of the laser endoscope apparatus according to Embodiment 1. FIG. 17C is a diagram for explaining the operation of the laser endoscope apparatus according to Embodiment 1. FIG. 17D is a diagram for explaining the operation of the laser endoscope apparatus according to the first embodiment. FIG. 17E is a diagram for explaining the operation of the laser endoscope apparatus according to the first embodiment. FIG. 18 is a flowchart showing an example of the operation of the laser endoscope apparatus. FIG. 19 is a schematic diagram for generating a panoramic image using the laser endoscope apparatus according to the first modification of the first embodiment. FIG. 20 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the second modification of the first embodiment. FIG. 21 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the third modification of the first embodiment. FIG. 22 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the fourth modification of the first embodiment. FIG. 23 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the fifth modification of the first embodiment. FIG. 24 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the sixth modification of the first embodiment. FIG. 25 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the seventh modification of the first embodiment. FIG. 26 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the eighth modification of the first embodiment. FIG. 27 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the ninth modification of the first embodiment. FIG. 28 is a schematic diagram of staining the inside of a living body using the laser endoscope apparatus according to Modification 10 of Embodiment 1. FIG. 29 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the eleventh modification of the first embodiment. FIG. 30 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the modification 12 of the first embodiment. FIG. 31 is a schematic diagram of imaging the inside of a living body using the laser endoscope apparatus according to the modification 12 of the first embodiment. FIG. 32A is a schematic diagram showing a state in which the inner wall of the digestive tract is imaged using a multiphoton laser microscope. FIG. 32B is a panoramic image showing the cell morphology at a position 50 μm deep from the inner wall surface (mucosal surface). FIG. 33 is a diagram showing an endoscope of the laser endoscope apparatus according to the second embodiment. FIG. 34A is a three-dimensional data image showing cell morphology at a depth within a predetermined range from the inner wall surface (mucosal surface), and is an image obtained by extracting a color region stained with both the curcumin dye and the acid red dye. FIG. 34B is a three-dimensional data image showing the cell morphology at a depth within a predetermined range from the inner wall surface (mucosal surface), and is an image obtained by extracting a color region stained with curcumin pigment. FIG. 34C is a three-dimensional data image showing cell morphology at a depth within a predetermined range from the inner wall surface (mucosal surface), and is an image obtained by extracting a color region stained with acid red pigment FIG. 35 is a block diagram showing a control configuration of the laser endoscope apparatus according to the third embodiment. FIG. 36A is an image showing a cell morphology at a position 50 μm deep from the inner wall surface (mucosal surface), and is an image obtained by extracting a color region stained with curcumin pigment. FIG. 36B is an image showing a cell morphology at a position 50 μm deep from the inner wall surface (mucosal surface), and is an image obtained by extracting a color region stained with acid red pigment. FIG. 36C is an image showing the cell morphology at a position 50 μm deep from the inner wall surface (mucosal surface), and is an image obtained by extracting color regions stained with both the curcumin dye and the acid red dye. FIG. 37 shows a group of cancer cells in the stomach stained with curcumin dye. FIG. 38 shows an image when a cell group inside a living body is imaged from a direction perpendicular to the inner wall surface (mucosal surface) using a confocal laser microscope. FIG. 39 shows an image when a cell group inside a living body is imaged from the upper right side with respect to the inner wall surface (mucosal surface) using a confocal laser microscope. FIG. 40 shows an image when a cell group inside a living body is imaged from the upper left side with respect to the inner wall surface (mucosal surface) using a confocal laser microscope. FIG. 41 shows an image when a cell group inside a living body is imaged from the upper right side with respect to the inner wall surface (mucosal surface) using a confocal laser microscope. FIG. 42 is an image obtained by imaging a living cell stained with a stain optimized in lysis method with a confocal laser microscope, (a) is a normal colonic mucosa, (b) is an image showing colon cancer. FIG. 43 is a schematic diagram showing an end portion on the distal end side of the endoscope of the laser endoscope apparatus according to the fourth embodiment. FIG. 44 is a schematic view showing the entire endoscope. FIG. 45 is a block diagram showing a control configuration of the laser endoscope apparatus FIG. 46 shows an image of an unstained colonic mucosa inner surface taken using a confocal laser microscope. FIG. 47 shows an image of an unstained colonic mucosa inner surface taken using a multiphoton laser microscope.

(Knowledge 1 as the basis of the present invention)
Of Knowledge 1, Knowledge 2, Knowledge 3, Knowledge 4, Knowledge 5, and Knowledge 6 that are the basis of the present invention, first, Knowledge 1 that is the basis of the present invention and the main configuration of the invention related to Knowledge 1 will be described. To do.

First, the relationship between the internal structure of a living body and cancer cells will be described.

The inside of a living body includes organs such as the digestive tract, respiratory organs, renal urinary organs, uterine ovarian genital organs, and cerebrospinal nerves. Examples of the digestive tract include the esophagus, stomach, small intestine, and large intestine.

FIG. 1 is a schematic diagram showing the arrangement of cells of the large intestine, which is an example of the digestive tract 112. For example, the inner wall of the large intestine is composed of a gland 130 that secretes mucus and an epithelium 120 that is in contact with food and absorbs water on the inner wall surface (mucosal surface) 113 side of the gland 130. The epithelium 120 is composed of a plurality of epithelial cells 121 arranged along the inner wall surface 113. The epithelial cell 121 has a nucleus 125 and a cytoplasm 126. The gland 130 has a shape in which a part of the epithelium 120 is recessed in a pot shape. The gland 130 is composed of a plurality of gland cells 131, and the gland cells 131 have a nucleus 135 and a cytoplasm 136. The portion where the gland 130 is depressed is called the crypt of the gland 130. A basement membrane 137, capillaries 132, and connective tissue 133 are formed inside the epithelial cells 121 and around the gland cells 131. A thin mucus layer secreted from the gland 130 is formed on the surface of the epithelial cell 121, and the epithelial cell 121 is protected by this mucus layer.

The size of the

nuclei

125 and 135 in the internal structure of the living body, the way in which the

nuclei

125 and 135 are arranged, and the distance from the nucleus 135 to the basement membrane 137 are important determination factors in making a pathological diagnosis such as cancer.

FIG. 2 is a diagram schematically showing a cancer cell population 152 that occurs in the digestive tract 112. It is said that the early stage cancer cell population 152 occurring in the gastrointestinal tract 112 is generally generated at a position within a depth of about 1 mm from the inner wall surface (mucosal surface) 113 of the gastrointestinal tract 112. If the cancer cell population 152 in the early stage, which is a state before reaching and exceeding the mucosal muscular plate 160, can be found over a wide area without omission, it expands beyond the mucosal muscular plate 160 and causes metastasis to other organs. This can reduce the number of cases that lead to advanced cancer.

At this time, as an attempt to grasp the lesion inside the living body including the cancer cell population 152, the inventors imaged the cell morphology inside the living body using a multiphoton laser microscope (FV1000MPE manufactured by Olympus). A multiphoton laser microscope is a fluorescence microscope using a multiphoton excitation process. A mouse was used as the living body.

FIG. 3 is a schematic diagram showing a state in which the inner wall of the digestive tract 112 is imaged using a multiphoton laser microscope. As shown in FIG. 3, the objective lens 16 of the multiphoton laser microscope is disposed to face the inner wall surface 113 of the digestive tract 112 in order to irradiate the inner wall of the digestive tract 112 that is an imaging target with the laser L.

When mainly imaging epithelial cells 121, the objective lens 16 is arranged so that the focal point of the objective lens 16 is tied to the inner wall surface (mucosal surface) 113. Thereby, the epithelial cells 121 and the like appear as shown in FIG. 3A, which is a schematic diagram cut along the aa line in FIG. Further, when mainly imaging the gland cells 131, the capillaries 132 and the connective tissue 133, the objective lens 16 is disposed so that the focal point of the objective lens 16 is deeper than the inner wall surface (mucosal surface) 113 by 10 μm or more. To do. As a result, the gland cells 131, the capillaries 132, and the connective tissue 133 appear as shown in FIG. 3B, which is a schematic diagram cut along the line bb in FIG. Thus, by changing the focal position of the objective lens 16 of the multiphoton laser microscope, the epithelial cells 121, gland cells 131, capillaries 132, and connective tissues 133 in the digestive tract 112 can be imaged.

In addition, when imaging the cell morphology inside a living body using a multiphoton laser microscope, the inventors dyed the living body (mouse) with a chromatic color using a dye containing an edible dye and performed imaging. . By using a staining agent, the epithelial cells 121, gland cells 131, capillaries 132, and connective tissues 133 of the digestive tract 112 can be selectively stained. In addition, an edible pigment | dye is a pigment | dye (for example, pigment | dye for food coloring) permitted administration to a human among natural pigment | dye or artificial synthetic pigment | dye.

The image shown in FIG. 4 is an image when epithelial cells 121 and gland cells 131 are imaged using a multiphoton laser microscope after staining epithelial cells 121 and gland cells 131 with a stain containing curcumin. The laser wavelength was 780 nm, and the magnification of the objective lens was 10 times for (a) and 25 times for (b). The image shown in FIG. 5 was obtained by imaging the capillary 132 and the connective tissue 133 using a multiphoton laser microscope after staining the capillary 132 and the connective tissue 133 with a stain containing Acid Red (Red 106). It is an image of the case. The laser wavelength was 840 nm, and the magnification of the objective lens was 10 times (a), 25 times (b), and 75 times (c) (25 times and 3 times zoom). As the staining agent containing curcumin, a curcumin solution (stock solution 5%) diluted 1/5 to 1/5000 with physiological saline can be used, and the staining time is about 2 to 5 minutes, respectively. FIG. 4 is black and white but is originally a color image, and by staining, the epithelium and glands can be separated into a green fluorescent color, and the capillaries and connective tissue can be separated into a dark green color with no fluorescent color. . This fluorescent color is expressed by correcting the actual fluorescence into a color that can be easily visually judged on the image.

As shown in FIGS. 4 and 5, when a cell group inside a living body is stained and imaged using a staining agent, a clear image of the cell group can be obtained.

In addition, as a new attempt, the inventors of the present invention have selected two or more chromatic colors that are different depending on the type of cells using two types of stains when staining a living body with a stain. And imaged. Specifically, the living body was stained using a stain containing curcumin and a stain containing acid red. Hereinafter, staining a cell group inside a living body with two staining agents into two or more selective chromatic colors depending on the cell type is referred to as “double staining”.

The image shown in FIG. 6 is obtained when the inner wall of the digestive tract 112 is double-stained with a stain containing curcumin and a stain containing acid red, and then the inner wall of the digestive tract 112 is imaged using a multiphoton laser microscope. It is an image. Among them, (a) is an image showing a normal gastrointestinal tract 112, and (b) is an image showing the gastrointestinal tract 112 in which a cancer cell population 152 of the colon at an early stage is formed. The magnification of the image shown in (a) is taken at a magnification of 1.5 times that of the image shown in (b).

In addition, as a staining agent containing curcumin, a curcumin solution (stock solution 5%) diluted 1/10 with physiological saline was used. The acid red solution (stock solution 10 mg / mL) was used as it was as a stain containing Acid Red. The staining time was 2 to 5 minutes each. Here, as a staining agent containing curcumin, a curcumin solution (stock solution 5%) diluted 1/5 to 1/5000 with physiological saline, and as a staining agent containing acid red, acid red solution (stock solution 10 mg / mL) ) Can be dyed even at a concentration of 1 to 1/1000 diluted as it is.

As shown in FIG. 6 (a), a plurality of tissues such as epithelium, glands, capillaries, or connective tissues in the inner wall of the digestive tract 112 are obtained by double-staining and then imaging the cells inside the living body. It can be confirmed simultaneously in one image. FIG. 6 is a black and white image, but is originally a color image. Due to the difference in staining tendency with the staining agent, the epithelium and glandular portions have a green fluorescent color, and the capillaries and connective tissue have a fluorescent color close to light red to orange. Compared with the case of staining with one staining agent, the epithelium and gland portions, capillaries and connective tissue can be separated and expressed more clearly. The fluorescent color resulting from the staining of the curcumin solution is displayed in green, the fluorescent color resulting from the staining of the acid red solution is displayed in red, and the actual fluorescence is expressed on the image with a color that is easy to visually determine. . In addition, as shown in FIG. 6B, the digestive tract 112 is in a normal state by comparing the shape and brightness of the cell groups represented in (a) and (b), or It can be confirmed whether lesions such as cancer occur.

In addition to the above, a staining agent containing curcumin and a staining agent containing Fast Green FCF (FastGreen FCF) can also be used as a staining agent for performing double staining. In this case, as a staining agent containing curcumin, a curcumin solution (stock solution 5%) diluted 1/10 with physiological saline was used, and as a staining agent containing fast green FCF, a fast green FCF solution (stock solution 10 mg / day) was used. mL) may be used as it is. The staining time may be 2 to 5 minutes. Here, as a staining agent containing curcumin, a curcumin solution (stock solution 5%) diluted 1/5 to 1/5000 with physiological saline, and as a staining agent containing Fast Green FCF, Fast Green FCF solution (stock solution 10 mg) / ML) can be stained even at a concentration of 1 to 1/1000 diluted as it is.

In addition, when dyeing a living body using a staining agent, the inventors specifically stain a cancer cell peripheral cell group other than cancer cells located around the cancer cell population 152 in a chromatic color. The cell group inside the living body was selectively stained using and the imaging was performed. Specifically, the living body was stained with a staining agent containing Rose Bengal.

(A) in FIG. 7 is an image obtained by imaging fluorescence emitted only from cancer cells using GFP, which is a green fluorescent protein. FIG. 7B is an image when the inner wall of the digestive tract 112 is imaged using a multiphoton laser microscope after staining the inner wall of the digestive tract 112 with a stain containing rose bengal. (C) in FIG. 7 is an image obtained by synthesizing (a) and (b). In an actual image, (a) is green fluorescent color and (b) is red fluorescent color. Only a group of cells can be identified.

As shown in (b) of FIG. 7, among the cell groups inside the living body, fluorescence can be obtained by the cancer cell peripheral cell groups located around the cancer cell population 152 shown in (a). It is possible to confirm whether cancer has occurred in the digestive tract 112 by imaging surrounding cell groups. Use of this image is effective in determining a treatment range for preventing recurrence after cancer cell removal treatment as well as cancer cells. For example, treatment is performed by determining criteria such as removing the cell group around the cancer cell shown in FIG. 7B from the cancer cell and the cell group around the cancer cell of half length from the cancer cell side. By doing, safer treatment for the patient can be performed.

Furthermore, the inventors tried to create a composite image by imaging the cell morphology inside the living body using a multi-photon laser microscope and superimposing a plurality of captured images.

FIG. 8 is a schematic diagram showing a state in which the inner wall of the digestive tract 112 is imaged using the multiphoton laser microscope 102 in a full circle. As shown in FIG. 8, after the imaging head 11 for irradiating the laser L is inserted into the expanded inner wall 113 of the digestive tract 112, the imaging head 11 is moved and imaging is performed in a plurality of imaging regions P. It can be carried out. At that time, it is possible to perform imaging so that adjacent imaging areas P1 and P2 among a plurality of imaging areas P overlap. Then, a composite image was created by superimposing areas Pa where adjacent imaging areas P1 and P2 overlap each other.

FIG. 9 is a composite image of the inner wall of the digestive tract 112 stained with a stain containing acid red. FIG. 10A is a composite image of the inner wall of the digestive tract 112 stained with a stain containing curcumin and a stain containing acid red. FIG. 10B is a diagram illustrating an example of a three-dimensional reconstruction of a panoramic image of the inner wall of the digestive tract 112.

As shown in FIG. 9 and FIG. 10A, by creating a composite image in which a plurality of images are superimposed, the cell morphology inside the living body can be grasped in a wide range and without omission. Further, as shown in FIG. 8, a panoramic image of the inner wall of the digestive tract 112 may be created by moving the imaging head 11 along the inner periphery of the digestive tract 112 and rotating 360 ° to capture images. Conceivable. Also, as shown in FIG. 10B, by reconstructing the panoramic image in a tunnel shape, it is easy to visualize and grasp at which position (coordinates) in the digestive tract 112 the lesion exists. By performing such imaging, it is possible to comprehensively detect lesions inside the living body.

That is, according to the main configuration of the present invention, a minute lesion such as a very early cancer (diameter 0.2 mm to 1 mm) which is too small to be noticed even by the current endoscope is not accidentally detected. It becomes possible to detect exhaustively over a wide range.

(Embodiment 1)
Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.

Note that each of the embodiments described below shows a preferred specific example of the present invention. The numerical values, shapes, materials, constituent elements, arrangement positions and connecting forms of the constituent elements, steps, order of steps, and the like shown in the following embodiments are merely examples, and are not intended to limit the present invention. The invention is specified by the claims. Therefore, among the constituent elements in the following embodiments, constituent elements not described in the independent claims are described as arbitrary constituent elements. Moreover, in each figure, the same code | symbol is attached | subjected to the substantially same structure, The overlapping description is abbreviate | omitted or simplified.

[1. Configuration of laser endoscope apparatus]
The laser endoscope apparatus according to the present embodiment is an apparatus that can image a lesion occurring in the digestive tract, respiratory tract, renal urinary tract, uterine ovarian genitalia, cerebrospinal nerve and the like over a wide range without leakage. In addition to imaging, it is possible to treat a lesion occurring in a living body. In the present embodiment, the digestive tract 112 inside the living body will be described as an example.

[1.1 Configuration for imaging preparation]
First, the configuration of a laser endoscope apparatus for preparing for imaging will be described.

Since the inner wall of the gastrointestinal tract 112 is actually uneven, it is desirable to push the gastrointestinal tract 112 into an imageable state before imaging using the laser endoscope device. In addition, in order to obtain a clear image using a laser endoscope apparatus, it is desirable to stain a cell group on the inner wall of the digestive tract 112 with a staining agent. Therefore, the laser endoscope apparatus according to the present embodiment includes an insertion tube that pushes the inside of the digestive tract 112, and a stain supply unit that supplies a stain to stain a group of cells on the inner wall of the digestive tract 112. ing.

(A) of FIG. 11 is a figure which shows the state after inserting the insertion tube 20 in the digestive tract 112. FIG.

As shown in FIG. 11A, the insertion tube 20 is provided with a supply port 42 for supplying fluid and a recovery port 43 for recovering the supplied fluid. The insertion tube 20 is provided with a first balloon 21 and a second balloon 22. The first balloon 21 and the second balloon 22 are expanded and contracted when fluid (gas or liquid) is taken in and out of the

balloons

21 and 22. The first balloon 21 is provided on the distal end side of the insertion tube 20 with respect to the supply port 42, and the second balloon 22 is provided on the rear side (opposite side of the distal end) with respect to the recovery port 43. By inflating the first balloon 21 and the second balloon 22 in the digestive tract 112, the space in the digestive tract 112 sandwiched between the first balloon 21 and the second balloon 22 becomes a closed space S.

FIG. 11B and FIG. 12 are diagrams illustrating an example of a staining agent supply unit 40 for supplying the staining agent 45. As shown in FIG. 12, for example, the staining agent 45 is supplied into the space S from the staining agent supply unit 40 in which the staining agent 45 is stored through the insertion tube 20 and the supply port 42. The staining agent 45 may be, for example, a staining agent containing curcumin or one acid red staining agent, but two types of staining agents including a staining agent containing curcumin and a staining agent containing acid red are used. Is desirable. By using two types of staining agents 45 and staining a cell group inside a living body in two colors, a clearer captured image can be obtained. Curcumins include not only curcumin but also curcuminoids (a mixture of several types of curcumin derivatives) with high water solubility.

The stain 45 may be one stain containing both curcumin and acid red. The stain 45 may be one stain containing both curcumin and fast green FCF, or two kinds of stains including a stain containing curcumin and a stain containing fast green FCF. There may be. Further, the staining agent 45 is not limited to two types, and may be one type of single color. For example, the staining agent 45 may be a staining agent including Rose Bengal. Moreover, before dyeing | staining, you may wash | clean the inside of the digestive tract 112 using the supply port 42 and the collection | recovery port 43, or remove mucus.

Thereafter, as shown in FIG. 13A, for example, gas is supplied from the supply port 42 to swell the inside of the digestive tract 112, whereby the inner wall of the digestive tract 112 is extended and flattened. The unevenness of the inner wall surface 113 when flattened is desirably such that the height difference between the concave and convex is within 0.2 mm, for example. Thereby, the preparation for imaging a living body using the laser endoscope apparatus is completed.

By flattening the inner wall of the digestive tract 112, it is possible to accurately grasp the state of the inner wall surface 113 and the cell group located at a predetermined depth from the inner wall surface 113. In addition, the inside of the digestive tract 112 is not inflated with a bag and imaged through the bag, but the laser L is directly applied to the inner wall of the digestive tract 112, so that the inner wall surface 113 and the like can be accurately grasped. it can.

The medium that swells in the digestive tract 112 is not limited to gas, and liquids such as distilled water and physiological saline can also be used. However, when a liquid is used, the liquid needs to transmit the wavelength of the laser to be used. When the medium is a liquid, it is desirable that the concentration of the staining agent be higher than that of a gas. In order to adjust the internal pressure of the digestive tract 112, a pressure sensor may be provided between the first balloon 21 and the second balloon. It is preferable to provide a plurality of pressure sensors at equal intervals.

[1.2 Basic configuration of laser endoscope apparatus]
Next, the basic configuration of the laser endoscope apparatus 1 according to the first embodiment will be described with reference to FIG. 13B, FIG. 14, and FIG.

FIG. 13B is a schematic diagram showing an end portion on the distal end side of the endoscope 2 of the laser endoscope apparatus 1 in FIG. FIG. 14 is a schematic diagram showing the entire endoscope 2. FIG. 15 is a block diagram showing a control configuration of the laser endoscope apparatus 1.

As shown in FIG. 15, the laser endoscope apparatus 1 includes an imaging unit 10 having an endoscope 2, a control unit 50, and an image processing unit 70. Further, the laser endoscope apparatus 1 includes a laser oscillator 60 and an optical component 65.

The laser L oscillated from the laser oscillator 60 is reflected by the dichroic mirror 66 that is the optical component 65, and further reflected by the mirror 19 in the endoscope 2 to irradiate the living body. The living cells irradiated with the laser L generate fluorescence, and the light due to the fluorescence is reflected by the mirror 19, passes through the dichroic mirror 66, and is detected by the photodetector 35. The light detected by the photodetector 35 is converted into an electrical signal, and an image is formed by the image processing unit 70. Since the fluorescence color varies depending on the staining agent, a plurality of photodetectors 35 are provided, and an optical filter for separating the colors can be placed in front of the photodetectors 35 to separate them.

As the laser oscillator 60, one having a pulse width of tens to hundreds of femtoseconds and a pulse frequency of tens to hundreds of MHz is used. The laser L in the present embodiment is a two-photon laser that is a kind of multi-photon laser, and the laser oscillator 60 uses, for example, a pulse laser having a wavelength of 800 nm and an output of up to 3.2 W. The laser output during imaging of this laser is emitted in the range of 0.16 to 0.32 W. By setting the wavelength to 800 nm or more, it is possible to prevent generation of photons in the ultraviolet region (wavelength of less than 400 nm) in half-wavelength light generated by the multiphoton excitation process. In the laser oscillator 60, the intensity of the laser L can be adjusted.

The dichroic mirror 66 that is the optical component 65 reflects the same wavelength as the laser L and transmits light of other wavelengths. Therefore, the laser L oscillated from the laser oscillator 60 is reflected toward the mirror 19 by the dichroic mirror 66. On the other hand, the fluorescence generated in the living cell is reflected by the mirror 19, passes through the dichroic mirror 66, and reaches the photodetector 35. The optical component 65 can also be configured by a prism, a 4 / λ plate, or the like.

The imaging unit 10 includes the endoscope 2 and the photodetector 35, and images the cell morphology inside the living body by applying a laser L to the inside of the living body.

The photodetector 35 detects the fluorescence generated by applying the laser L, and converts the fluorescence into an electrical signal corresponding to the fluorescence intensity. As the photodetector 35, for example, a photomultiplier tube, a CCD semiconductor image sensor, or the like can be used.

The endoscope 2 includes an inner cylinder 12 and an outer cylinder 13 that surrounds a part of the inner cylinder 12 as shown in FIG. A part of the inner cylinder 12 and the outer cylinder 13 are inserted into the living body. The length of the inner cylinder 12 is, for example, 50 mm, and the outer diameter of the inner cylinder 12 is, for example, 3 to 10 mm. A linear actuator is attached to the inner cylinder 12, and the inner cylinder 12 can move about 25 mm in the axial direction X with respect to the outer cylinder 13. The inner cylinder 12 is equipped with an ultrasonic motor, and the inner cylinder 12 can rotate 360 ° with respect to the outer cylinder 13. The operation of the inner cylinder 12 in the axial direction X or the operation in the rotation direction R is controlled by the control unit 50.

An imaging head 11 is provided at the end of the endoscope 2 on the distal end side of the inner cylinder 12. As shown in FIG. 13B, the imaging head 11 passes through the side of the insertion tube 20 and is inserted into the living body together with the inner cylinder 12. The imaging head 11 is controlled to move inside the living body by operations in the axial direction X and the rotation direction R of the inner cylinder 12.

The imaging head 11 has an objective lens 16, a focus variable unit 18, a spacer 17, and a mirror 19.

As described above, the mirror 19 changes the direction of the laser L output from the laser oscillator 60 toward the objective lens 16 or changes the direction of the light fluorescent by the living cells toward the photodetector 35. It is.

The objective lens 16 is provided to face the inner wall surface 113 of the living body. The objective lens 16 has, for example, a diameter of 10 mm, a magnification of 10 times, a resolution of 5 μm, and an imaging field of view of 3 mm × 3 mm. Alternatively, the objective lens 16 has a diameter of 12 mm, a magnification of 40 times, a resolution of 10 μm, and a visual field of 7.5 mm × 7.5 mm. The wider the field of view, the better. The objective lens 16 may be a lens having a diameter of 3 mm to 5 mm that can be easily inserted into a living body by cutting a part of the lens having the diameter described on the left or an objective lens that can obtain the same resolution.

The focus variable unit 18 is, for example, a piezoelectric actuator or an electromagnetic actuator, and changes the focus position of the objective lens 16 by moving the objective lens 16 in the direction of the optical axis. The focus variable unit 18 is controlled in operation by the control unit 50 so that the focus can be adjusted from the inner wall surface (mucosal surface) 113 within a depth range of 0 to 1000 μm. By changing the focal position, the state of the living body at a predetermined depth from the cell surface of the digestive tract 112 can be imaged. When a confocal laser is used instead of the multiphoton laser, the depth of the focal point from the inner wall surface (mucosal surface) 113 may be adjusted in the range of 0 to 75 μm.

The spacer 17 has, for example, a ring shape and is provided around the space between the objective lens 16 and the inner wall surface 113. The spacer 17 is a component for preventing the objective lens 16 from touching the inner wall of the living body and for maintaining the distance between the objective lens 16 and the inner wall surface 113 constant. The distance between the objective lens 16 and the inner wall surface (mucosal surface) 113 can be appropriately set within a range of, for example, 1 mm or more and 10 mm or less by replacing the spacer 17 before starting imaging or adding a mechanism that can be changed by an actuator or the like. Set to a value. The control unit 50 controls the movement of the imaging head 11 (inner cylinder 12) while bringing the spacer 17 into contact with the inner wall surface 113, and maintains the distance of the objective lens 16 with respect to the inner wall surface 113 constant.

The control unit 50 includes a CPU, a ROM, a RAM, and the like. The control unit 50 controls the operation of the imaging head 11 via the inner cylinder 12. Specifically, the control unit 50 controls the imaging head 11 to move in the circumferential direction along the inner circumference of the inner wall of the digestive tract 112, and in the duct direction (digestive tract axis) of the digestive tract 112. Move control along. In addition, the control unit 50 controls the operation of the focus changing unit 18 to change the position of the objective lens 16 in the optical axis direction and to control the focal position connected to the inside of the living body. The control unit 50 can also adjust the laser output by controlling the laser oscillator 60.

The image processing unit 70 stores the electrical signal (fluorescence intensity) converted by the photodetector 35 and the coordinate position of the imaging unit 10 sent from the control unit 50 in association with each other, and processes these data to perform digital processing. Generate an image. The generated digital image is displayed on a monitor, printed out, or recorded in a storage device, for example. As an example of the coordinate position of the imaging unit 10, a distance from a location (for example, throat or anus) serving as a reference for the patient, a rotation angle of the imaging head 11, and the like can be used.

In the laser endoscope apparatus 1 according to the present embodiment, the control unit 50 controls the movement of the imaging head 11 so that the imaging head 11 scans with a constant distance from the inner wall surface 113 of the living body. . Then, as illustrated in FIG. 16, the imaging unit 10 captures a plurality of imaging regions P that are imaged with the movement of the imaging head 11 such that adjacent imaging regions P1 and P2 partially overlap each other. The image processing unit 70 generates a composite image by superimposing areas Pa where adjacent imaging areas P1 and P2 overlap each other. Thereby, the cell form inside the living body can be imaged over a wide range without leakage.

Further, a panoramic image can be generated by using the laser endoscope apparatus 1. For example, as shown in FIG. 16, the control unit 50 controls the imaging head 11 to move 360 ° along the inner periphery of the digestive tract 112 (or to revolve around the axis of the digestive tract 112). And the imaging part 10 images the some imaging area P imaged with the movement of the imaging head 11 so that a part of adjacent imaging area P1 and P2 may overlap in the circumferential direction. The image processing unit 70 generates a panoramic image by superimposing areas Pa where adjacent imaging areas P1 and P2 overlap each other. Thereby, the state of the inner wall of the digestive tract 112 can be comprehensively grasped.

Also, by using this laser endoscope apparatus 1, an image along the duct direction of the digestive tract 112 can be generated. For example, as shown in FIG. 16, after the imaging at the inner circumference 360 ° in the digestive tract 112 is finished, the control unit 50 moves the imaging head 11 by a predetermined distance along the duct direction of the digestive tract 112, The imaging unit 10 images the moved imaging area P11 so as to partially overlap the imaging area P1 adjacent in the duct direction. Then, the image processing unit 70 superimposes the region Pb where the two imaging regions P1 and P11 overlap each other. Next, the imaging unit 10 again performs imaging at the inner circumference 360 ° of the digestive tract 112, and the image processing unit 70 superimposes the overlapping areas in the circumferential direction and the pipeline direction and extends in the pipeline direction. Generate a panoramic image. According to this, it is possible to comprehensively grasp the state of the inner wall of the digestive tract 112 also in the duct direction.

Also, by using this laser endoscope apparatus 1, a stereoscopic image of a living body can be generated. For example, the control unit 50 controls the focus variable unit 18 of the imaging head 11 to change the focal position of the objective lens 16, and the imaging unit 10 performs a plurality of imaging operations with different depths as the focal position changes. Image the area. And the image processing part 70 produces | generates the three-dimensional image of the cell form inside a biological body by arrange | positioning the several image obtained by imaging corresponding to a focus position. Thereby, not only the inner wall surface 113 of the living body but also the cell form inside the living body having a depth within a predetermined range can be imaged.

[2.1 Operation of laser endoscope apparatus 1]
Next, the operation of the laser endoscope apparatus 1 when imaging the cell morphology inside the digestive tract 112 will be described.

As shown in FIG. 17A (a), since the inner wall of the digestive tract 112 is normally in an uneven state, an operation for flattening the inner wall of the digestive tract 112 is performed.

First, as shown in FIG. 17A (b), the insertion tube 20 is inserted into the digestive tract 112.

Next, as shown in FIG. 17A (c), the first balloon 21 is expanded and brought into contact with the inner wall surface 113 of the digestive tract 112 at the distal end of the insertion tube 20. In addition, in order to improve airtightness, the 1st balloon 21 is comprised by three balloons.

Next, as shown in FIG. 17A (d), the second balloon 22 located behind the insertion tube 20 is expanded and brought into contact with the inner wall surface 113 of the digestive tract 112. The second balloon 22 is also composed of three balloons. By this operation, a closed space S is formed between the first balloon 21 and the second balloon 22. Then, air is blown out from the supply port 42 into the closed space S, and the inside of the digestive tract 112 is expanded. Thereby, the wrinkles etc. which exist in the inner wall of the digestive tract 112 are extended and planarized. In addition, you may perform the process of planarizing an inner wall after dyeing mentioned later.

Next, as shown in FIG. 17B (e), the cleaning liquid is supplied from the supply port 42 to the closed space S. Thereby, the inner wall surface 113 of the digestive tract 112 is washed. Thereafter, the cleaning liquid is sucked from the recovery port 43 and recovered.

Next, as shown in FIG. 17B (f), the pronase solution is supplied from the supply port 42 to the closed space S. Thereby, excess mucus attached to the inner wall surface 113 of the digestive tract 112 is removed. Thereafter, the pronase solution is sucked through the collection port 43 and collected.

Next, as shown in FIG. 17B (g), a stain A (for example, a stain containing curcumin) is supplied from the supply port 42 to the closed space S and filled. Then, after standing for 2 to 5 minutes, wash with a cleaning solution. Thereby, a predetermined cell group on the inner wall of the digestive tract 112 is stained with the stain A. Note that the predetermined cell group indicates a plurality of cells included in the epithelial cells 121, the gland cells 131, the capillaries 132, the connective tissue 133, and the like.

Next, as shown in FIG. 17B (h), a stain B (for example, a stain containing acid red) is supplied to the space S closed from the supply port 42 and filled. Then, after standing for 2 to 5 minutes, wash with a cleaning solution. Thereby, a predetermined cell group on the inner wall of the digestive tract 112 is stained with the stain B, and the inner wall of the digestive tract 112 is double-stained. As described above, if the method of filling the space S with the stain A or B is used, the inner wall of the digestive tract 112 can be double-stained with little unevenness.

Next, as shown in (i) of FIG. 17C, the endoscope 2 is inserted into the closed space S. The imaging head 11 provided on the distal end side of the endoscope 2 keeps the distance between the inner wall surface 113 and the objective lens 16 constant, and makes the focal position of the objective lens 16 deep from the inner wall surface (mucosal surface) 113. Imaging is performed in accordance with 0 μm, 30 μm, 60 μm, 90 μm, 120 μm, and 150 μm. Note that the position at a depth of 0 μm can be determined by the autofocus function of the endoscope 2. When a confocal laser is used instead of the multiphoton laser, imaging is performed by adjusting the focal position of the objective lens 16 to positions of 0 μm, 25 μm, 50 μm, and 75 μm deep from the inner wall surface (mucosal surface) 113. The depth changing pitch is an example, and may be a finer pitch or a coarse pitch.

Then, as shown in FIG. 17C (j), imaging is performed while the imaging head 11 is rotated 360 ° along the inner wall surface 113. At this time, imaging is performed so that a part of the imaging regions P1 and P2 adjacent in the circumferential direction overlap each other. A first panoramic image is acquired by this partially overlapping imaging. When the imaging at the inner circumference of 360 ° is completed, the imaging head 11 is moved a predetermined distance along the duct direction of the digestive tract 112. Then, again, the imaging head 11 is rotated 360 ° to obtain a second panoramic image. Note that the first panorama image and the second panorama image are captured after setting the imaging regions P1 and P11 adjacent to each other in the duct direction of the insertion tube 20 to overlap each other. The pattern of overlapping portions in the circumferential direction and the straight line direction of the digestive tract is accurately combined by image processing to be combined into a seamless image. These operations are repeated a plurality of times (in this embodiment, 5 times).

Once these imaging operations are completed, the endoscope 2 including the imaging head 11 is moved rearward from the second balloon 22 as shown in (k) of FIG. 17C.

Next, the first balloon 21 is deflated as shown in (l) of FIG. Then, as shown in FIG. 17D (m), the insertion tube 20 is pulled backward while the position of the second balloon 22 is maintained. Thereafter, as shown in FIG. 17D (n), the first balloon 21 is expanded. After the second balloon 22 is deflated as shown in (o) of FIG. 17E, the insertion tube 20 is moved backward while maintaining the position of the first balloon 21 as shown in (p) of FIG. 17E. Pull. Thereafter, as shown in (q) of FIG. 17E, the second balloon 22 is expanded. Thereby, another closed space S1 adjacent to the previously formed closed space S in the pipe line direction is formed. Then, the operation shown in (d) of FIG. 17A to (k) of FIG. 17C is performed again in the closed space S1.

By repeating these operations, for example, imaging with a length of 300 mm in the pipe line direction can be performed. If the digestive tract 112 is the large intestine, the entire length of the large intestine may be divided into four images.

According to the operation of the laser endoscope apparatus 1 as described above, the state of the inner wall can be comprehensively and efficiently imaged in the inner circumferential direction and the duct direction of the digestive tract 112.

[2.2 Operation 2 of laser endoscope apparatus]
The laser endoscope apparatus 1 is configured to perform an operation for efficiently detecting a lesion such as a cancer cell in the depth direction.

Therefore, the controller 50 has the following two variable focus modes (see FIG. 15). Specifically, the control unit 50 changes the focus position at the first focus variable mode 51 that changes the focus position at the first pitch, and at the second pitch that is smaller than the first pitch. 2 focus variable modes 52. The first variable focus mode 51 is a mode in which, for example, the focal point is changed from the inner wall surface (mucosal surface) 113 of the living body to focus at depths of 0 μm, 30 μm, 60 μm, 90 μm, and 120 μm. The second variable focus mode 52 is a mode in which the focus position is changed at a finer pitch, for example, a pitch of 5 μm. These variable pitches can be changed by changing the program.

In addition, the control unit 50 stores in advance a reference image of normal cells in a state where there is no lesion for each corresponding internal organ in order to quickly determine the presence or absence of the lesion. The reference image of normal cells differs depending on the type of laser to be irradiated (multiphoton laser or confocal laser) and the depth from the cell membrane surface of the organ, and if a stain is used, it also depends on the type of stain It is preferable to prepare a reference image corresponding to the imaging conditions in advance.

FIG. 18 is a flowchart showing an example of the operation of the laser endoscope apparatus 1. First, the control unit 50 causes the first variable focus mode 51 having a rough pitch to perform one-way imaging in the depth direction (S11). At this time, the magnification is set to a low value (for example, 10 times or less), and it is determined based on the image at the time of trial shooting. good.

Next, the control unit 50 compares the image obtained in the first variable focus mode 51 with the reference image of the normal cell stored in advance with respect to at least one of shape and brightness, and determines the suspected lesion. (S12). If there is no suspicion of the lesion, the examination is terminated (S13).

When the control unit 50 determines that a portion suspected of having a lesion exists in the image obtained by imaging, the second focus is in the vicinity of the focus position when the image of the portion suspected of having a lesion is captured. Imaging is performed in the variable mode 52 (S14). At this time, the magnification is set high (for example, 40 times), and it is only necessary to pick up an image that can be diagnosed by imaging only at a certain depth without changing the depth or imaging of the surface.

Next, the control unit 50 compares the image obtained in the first variable focus mode 51 with the reference image of the normal cell stored in advance with respect to at least one of shape and brightness, and determines the suspected lesion. (S15). If there is no suspicion of a lesion, the examination is terminated (S16).

Then, when there is a diseased cell in the image obtained by the imaging unit 10, the control unit 50 increases the output of the laser L compared to the time of imaging, and increases the output to the diseased cell. The lesioned cells are removed (transpiration) (S17). Although not shown in the figure, after removing the diseased cells, it is preferable to image the same location again and reconfirm the presence or absence of the diseased cells. The laser output at the time of cell removal is 10 to 20 times that at the time of imaging and is 2 to 3 W.

The above determination may be made automatically by comparing the shape, brightness, etc. using a computer. In addition, it is preferable to make a doctor's judgment on a place where a lesion is suspected after the judgment by the computer.

By performing such tomographic imaging, comprehensive imaging can be performed while shortening the imaging time. In addition, the diseased cells can be removed early and reliably. In the above removal treatment, diagnosis and removal may be performed before moving in the imaging space S described with reference to FIGS. 17A to 17E, or image coordinates may be taken at another opportunity after imaging of the target organ is completed. May be used to form a series of imaging spaces S to remove the lesion. These can be determined by the patient's physical strength, the symptoms of the affected area, the performance of the laser endoscope apparatus 1, and the like.

(Modification 1)
FIG. 19 is a schematic diagram illustrating an example of generating a panoramic image using the laser endoscope apparatus 1 according to the first modification of the first embodiment.

The control unit 50 according to the first modification controls the imaging head 11 attached to the arm 15 so as to rotate spirally around the axis of the digestive tract 112. The imaging unit 10 images a plurality of imaging regions P to be imaged with the rotation of the imaging head 11 so that adjacent imaging regions P1 and P2 partially overlap in the rotation direction R. The image processing unit 70 generates a panoramic image by superimposing areas Pa where adjacent imaging areas P1 and P2 overlap each other. Thereby, the state of the inner wall of the digestive tract 112 can be comprehensively imaged.

(Modification 2)
FIG. 20 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the second modification of the first embodiment.

In the second modification, a pair of wheels 17a is attached to the arm 15 on the distal end side of the endoscope 2 in place of the spacer 17 in order to keep the distance between the inner wall surface 113 of the digestive tract 112 and the objective lens 16 constant. ing. When the imaging head 11 is moved in the inner circumferential direction, the objective lens 16 is kept at a certain distance from the inner wall surface 113 by rolling while the pair of wheels 17a are in contact with the inner wall surface 113. It can be moved. According to this, it is possible to accurately focus on the imaging target such as the epithelial cell 121, the gland cell 131, the capillary vessel 132, or the connective tissue 133.

(Modification 3)
FIG. 21 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the third modification of the first embodiment.

In Modification 3, a pressing member 23 is provided on the back of the imaging head 11 (on the side opposite to the laser L irradiation side). Then, the spacer 17 is brought into contact with the inner wall on the imaging region side by pressing the inner wall on the opposite side of the imaging region with the pressing member 23 by inflating the fluid into the pressing member 23. As a result, the distance between the objective lens 16 and the inner wall surface 113 can be maintained constant, and focusing with respect to the imaging target can be accurately performed.

(Modification 4)
FIG. 22 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the fourth modification of the first embodiment.

In Modification 4, instead of the pressing member 23 shown in Modification 3, a support roller 24 is provided on the back of the imaging head 11. Then, the spacer 17 is brought into contact with the inner wall on the imaging region side by pressing the support roller 24 against the inner wall on the opposite side of the imaging region using the expansion mechanism 25. As a result, the distance between the objective lens 16 and the inner wall surface 113 can be maintained constant, and focusing with respect to the imaging target can be accurately performed.

(Modification 5)
FIG. 23 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the fifth modification of the first embodiment.

In Modification 5, instead of the support roller 24 shown in Modification 4, a sliding member 26 is provided on the back of the imaging head 11. Then, the spacer 17 is brought into contact with the inner wall on the imaging region side by pressing the sliding member 26 against the inner wall on the opposite side of the imaging region using the expansion mechanism 25. As a result, the distance between the objective lens 16 and the inner wall surface 113 can be maintained constant, and focusing with respect to the imaging target can be accurately performed.

(Modification 6)
FIG. 24 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the sixth modification of the first embodiment.

In Modification 6, as shown in FIG. 24A, the imaging head 11 is supported by a joint mechanism 27 that moves flexibly. Further, another joint mechanism 28 is provided on the back of the imaging head 11 to support the imaging head 11. According to this structure, as shown in FIG. 24 (b), it is also possible to image an uneven portion 113a such as a half-moon fold of the large intestine existing in the digestive tract 112. Further, by using the autofocus function, the distance between the objective lens 16 and the inner wall surface 113 can be kept constant.

(Modification 7)
FIG. 25 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the seventh modification of the first embodiment.

In Modification 7, the distal end of the endoscope 2 is inserted into the first balloon 21, and the imaging head 11 is provided in the middle of the inner tube 12 of the endoscope 2 (between the first balloon 21 and the second balloon 22). The joint mechanism 27 is supported. According to this, the joint mechanism 28 shown in the modification 6 is not necessary, and the structure of the endoscope 2 can be simplified as compared with the modification 6.

(Modification 8)
FIG. 26 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the eighth modification of the first embodiment.

In Modification 8, a gyro sensor 29 is provided in the imaging head 11. Thereby, the information regarding the position and orientation of the imaging head 11 at the time of imaging can be acquired. It is also possible to display the captured image data in 3D. In addition, according to this structure, after imaging with a coarse pitch using the first focus variable mode 51 described above, it quickly and accurately returns to a portion suspected of having a lesion, and the first pitch with a fine pitch is obtained. Imaging can be performed in the two variable focus modes 52. A GPS function may be added instead of the gyro sensor 29.

Further, in the modified example 8, the pressure sensor 30 is provided in the inner tube 12 of the endoscope 2. By measuring and feeding back the pressure in the closed space S using the pressure sensor 30, the pressure in the space S can be adjusted appropriately.

(Modification 9)
FIG. 27 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the modification 9 of the first embodiment.

In Modification 9, the imaging head 11 is provided with an extendable spacer 31 for adjusting the distance between the objective lens 16 and the inner wall surface 113. FIG. 27A shows a state where the expandable spacer 31 is expanded, and FIG. 27B shows a state where the expandable spacer 31 is contracted. The distance between the objective lens 16 and the inner wall surface 113 can be accurately adjusted by the expansion / contraction of the expansion / contraction spacer 31. The telescopic spacer 31 can be configured by an actuator or the like.

(Modification 10)
FIG. 28 is a schematic diagram illustrating an example of staining the inside of a living body using the laser endoscope apparatus 1 according to the tenth modification of the first embodiment.

In the tenth modification, the insertion tube 20 is provided with a plurality of discharge ports 42a. Then, the inner wall is dyed by spraying a staining agent from the discharge port 42a toward the inner wall. According to this, compared with the method of filling the space S with the stain, the amount of the stain used can be reduced. Therefore, even a staining agent using a dye having a small allowable amount can be used with confidence.

(Modification 11)
FIG. 29 is a schematic diagram illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the eleventh modification of the first embodiment.

In the modification 11, the distance between the pair of wheels 17a of the imaging head 11 shown in the modification 2 is widened so that the mirror 19 and the objective lens 16 can be moved in the axial direction X of the endoscope 2 by an actuator or the like (not shown). It is configured. According to this, the inside of the living body can be imaged without moving the inner cylinder 12 of the endoscope 2 more than necessary.

(Modification 12)
30 and 31 are schematic diagrams illustrating an example of imaging the inside of a living body using the laser endoscope apparatus 1 according to the first modification of the first embodiment.

In the modified example 12, the interval between the pair of wheels 17a of the imaging head 11 shown in the modified example 2 is widened, and a plurality of mirrors 19 and objective lenses 16 (five sets in this modified example) are provided between the pair of wheels 17a. Yes. And as shown to (a) and (b) of FIG. 31, the imaging head 11 is comprised so that the area | region between the 1st balloon 21 and the 2nd balloon 22 can be imaged by one 360 degree rotation operation | movement. Yes. According to this, imaging inside a living body can be performed efficiently.

In the above-described embodiment, the endoscope 2, the insertion tube 20, the inner cylinder 12, the outer cylinder 13 and the like are shown in a linear form, but in order to smoothly insert along the shape of the large intestine or the like. The endoscope 2, the insertion tube 20, the inner cylinder 12, the outer cylinder 13 and the like are preferably flexible, and an optical fiber or the like is preferably used as a laser waveguide. Further, in order to make the arm 15 or the like of the imaging head 11 into an L-shaped structure or to be linearly accommodated in the inner cylinder 12, it is fixed to an L-shape or the like with an appropriate joint structure and a wire or the like in the imaging head 11 or the like. This is possible by having a structure.

The endoscope 2, insertion tube 20, arm 15, spacer 17, balloons 21 and 22, etc. are made of metal, resin, rubber, etc., but directly touch a living organ such as the large intestine and stomach. For this reason, careful attention is paid to the surface processing, and the finishing is extremely accurate.

(Knowledge 2 as the basis of the present invention)
Next, Knowledge 2 that is the basis of the present invention and the main configuration of the invention related to Knowledge 2 will be described.

Knowledge 2 describes an example in which a multi-photon laser microscope (FV1000MPE manufactured by Olympus Corporation) is used to capture an internal cell form of a living body, and a panoramic image is created by superimposing a plurality of captured images. A mouse was used as the living body.

FIG. 32A is a schematic diagram showing a state in which the inner wall of the digestive tract 112 is imaged using the multiphoton laser microscope 102 in a full circle. The method for creating a panoramic image is almost the same as the method for creating FIG. 10A of Knowledge 1, and the imaging head 11 is moved along the inner periphery of the digestive tract 112 and rotated 360 ° for imaging. A panoramic image was obtained by combining the images.

FIG. 32B is a panoramic image showing the cell morphology at a depth of 50 μm from the inner wall surface (mucosal surface) 113. As a staining agent for staining a cell group, both a staining agent containing curcumin and a staining agent containing Acid Red (Red No. 106) were used. From FIG. 32B, it can be seen that a plurality of crypts 138 (or glands 130) are arranged at approximately regular intervals.

In FIG. 32B, the imaging regions P are arranged corresponding to the direction of the short hand of the watch shown in FIG. 32A, but an isolated lymph nodule in which a plurality of lymphocytes are gathered is formed in the direction of 8 o'clock. . The isolated lymph nodule is not currently worthy of a lesion such as cancer, but it can be seen that the crypt 138 of the gland 130 disappears in the region where the isolated lymph nodule is formed. Therefore, the inventors considered that the position of a lesion can be specified when a lesion is found by using the region where the isolated lymph nodule is formed as a marker of coordinates in the panoramic image. Moreover, even if there was no isolated lymph nodule, we considered that the position where the lesion exists can be clearly identified by using the predetermined position in the panoramic image as a reference. Hereinafter, an embodiment based on Knowledge 2 will be described.

(Embodiment 2)
The configuration of the laser endoscope apparatus 1A according to the second embodiment will be described with reference to FIG. FIG. 33 is a diagram showing the endoscope 2 of the laser endoscope apparatus 1A.

The endoscope 2 of the laser endoscope apparatus 1 </ b> A has substantially the same configuration as the endoscope 2 shown in the first embodiment, and includes an inner cylinder 12 and an outer cylinder 13 that surrounds a part of the inner cylinder 12. And. A linear actuator is attached to the inner cylinder 12, and the inner cylinder 12 is movable in the axial direction X with respect to the outer cylinder 13. The inner cylinder 12 is equipped with an ultrasonic motor, and the inner cylinder 12 can rotate 360 ° with respect to the outer cylinder 13. The operation of the inner cylinder 12 in the axial direction X or the operation in the rotation direction R is controlled by the control unit 50.

Furthermore, the endoscope of the laser endoscope apparatus 1A according to the present embodiment detects an angle detector 81 that detects an angle of the inner cylinder 12 in the rotation direction R, and a position of the inner cylinder 12 in the axial direction X. A linear scale 82 is provided. Since the endoscope 2 includes the angle detector 81 and the linear scale 82, for example, the distance in the axial direction X from the isolated lymph nodule to the lesion and the angle in the rotation direction R are determined with reference to the position of the isolated lymph nodule. And know the location of the lesion. Further, even if there is no isolated lymph nodule, it is possible to characterize a position where a lesion exists by using a predetermined position as a reference, such as an anus for the large intestine and a mouth for the stomach.

As described above, according to the laser endoscope apparatus 1A, a coordinate reference can be provided in the panoramic image, and it is possible to visualize and grasp at which position in the digestive tract 112 the lesion exists. In addition, by providing a coordinate reference, it is possible to show evidence captured by rotating 360 °, and to present to the patient that the acquired image is a non-leakage image having a full circumference.

(Knowledge 3 on which the present invention was based)
Next, Knowledge 3 that is the basis of the present invention and the main configuration of the invention related to Knowledge 3 will be described.

In Knowledge 3, a cell shape inside a living body is imaged by changing the depth of focus using a multiphoton laser microscope (Olympus FV1000MPE), and a plurality of captured images are cut at predetermined positions to obtain a cross-sectional image. An example of creating (tomographic image) will be described. A mouse was used as the living body.

34A, 34B, and 34C are images showing cell morphology at a depth within a predetermined range from the inner wall surface (mucosal surface), from the mucosal surface (depth 0) to a depth of 150 μm at a depth of 2 μm. This is a three-dimensional data image obtained by capturing and combining a total of 75 images. In each of FIGS. 34A to 34C, (a) is an image obtained by planarly viewing a cell group from a direction perpendicular to the inner wall surface 113, and (b) is a cross-sectional image when (a) is cut along the line bb. (C) is a cross-sectional image when (a) is cut along the line cc.

As the staining agent for staining the cell group, both staining agents including curcumin and staining agent including acid red (red No. 106) were used. The staining time was 5 minutes. The staining time is a period of time when the staining agent is brought into contact with the cell group and the staining dye is permeated between the cells themselves or each cell.

34A to 34C are images in which the same cell group is simultaneously imaged and different colors (wavelengths) are extracted by filtering. FIG. 34A is an image obtained by extracting color regions stained with both the curcumin dye and the acid red dye. FIG. 34B is an image obtained by extracting a color region stained with a curcumin pigment. FIG. 34C is an image obtained by extracting the color region stained with the acid red dye. FIGS. 34A to 34C are black and white, but are originally color images. Due to the difference in dyeing tendency due to the staining agent, the area stained with curcumin dye is green fluorescent color, and the area stained with acid red dye is light red. It is represented by a fluorescent color close to orange, and the color difference is more clearly represented.

34A to 34C show cancer tissue and normal mucosal tissue, and it can be seen that there is a difference in permeability depending on the respective pigments.

As shown in FIG. 34B, the curcumin pigment shows higher permeability in cancer tissues than in normal mucosal tissues. Specifically, in the case of curcumin pigment, the depth of staining is about 40 μm inside the cancer tissue, whereas it is about 20 μm inside the normal mucosal tissue.

As shown in FIG. 34C, Acid Red pigment shows lower permeability in cancer tissue than normal mucosal tissue. Specifically, in the case of Acid Red dye, the depth of staining is about 40 μm inside the cancer tissue, while it is about 70 μm inside the normal mucosal tissue.

Thus, there is a difference in the permeability of the pigment depending on whether the cell morphology is cancer tissue or normal mucosal tissue. Using this property, we can determine whether the cell group is a normal cell group or a cancer cell group by measuring the depth at which the cell group shown in the cross-sectional image is stained. It is done. Hereinafter, Embodiment 3 based on Knowledge 3 will be described.

(Embodiment 3)
The configuration of the laser endoscope apparatus 1B according to Embodiment 3 will be described with reference to FIG. FIG. 35 is a block diagram showing a control configuration of the laser endoscope apparatus 1B.

The laser endoscope apparatus 1B has substantially the same configuration as the laser endoscope apparatus 1 described in the first embodiment, and includes an imaging unit 10 having the endoscope 2, a control unit 50, an image processing unit 70, and a laser oscillator. 60 and an optical component 65.

Further, the laser endoscope apparatus 1B includes a stain supply unit 40 that supplies the stain to the inside of the living body (see FIG. 12). In the present embodiment, double staining is used in which a group of cells inside a living body is dyed into two or more selective chromatic colors that differ depending on the cell type.

The imaging unit 10 applies a laser to the stained cell group, changes the focal position (for example, 0 to 1000 μm), and images a plurality of imaging regions P having different depths. The image processing unit 70 creates a stereoscopic image by arranging a plurality of images obtained by imaging by the imaging unit 10 in correspondence with the focal position. Then, the stereoscopic image is cut at a predetermined position on the image including the stained cell group, thereby generating a cross-sectional image of the stained cell group.

The control unit 50 determines a suspicious lesion based on the depth at which the cell group represented in the cross-sectional image is stained. For example, if the depth at which the cell group is stained with the curcumin dye is larger than that of normal mucosal tissue (for example, 1.5 times or more), it is determined that cancer cells are generated, and if they are equivalent (for example, 1 (Less than 5 times) Judge that no cancer cells have developed. Further, if the depth at which the cell group is stained with the acid red dye is smaller than that of normal mucosal tissue (for example, less than 0.6 times), it is determined that cancer cells are generated, and if they are equivalent (for example, (0.6 times or more) Judge that cancer cells have not developed. In addition, after determining the presence or absence of cancer cells by single color or double staining or the like, the above-described cross-sectional image determination can be further improved.

The laser endoscope apparatus 1B according to the present embodiment includes an imaging head 11 that is inserted into a living body, an imaging unit 10 that captures an image of the living body by applying a laser to the living body via the imaging head 11, and imaging. A control unit 50 that controls the operation of the head 11 and an image processing unit 70 that processes an image captured by the imaging unit 10 are provided. The imaging head 11 includes an objective lens 16 and a focus variable unit 18 that can change the focal position of the objective lens 16 in the depth direction of the living body. The control unit 50 operates the focus varying unit 18 to change the focal position, and the imaging unit 10 images a plurality of imaging regions P having different depths as the focal position is changed. Then, the image processing unit 70 generates a cross-sectional image inside the living body by cutting a plurality of images obtained by imaging by the imaging unit 10 at a predetermined position.

It is possible to determine the presence or absence of canceration even in the depth direction inside the living body by generating a cross-sectional image with the laser endoscope apparatus 1B. In the case where examination and diagnosis are performed at different occasions, the image processing unit only needs to display an image that is currently captured, and by leaving other devices to generate panoramic images and stereoscopic images, The burden can be reduced.

(Findings 4, 5, and 6 that are the basis of the present invention)
Next, the main structures of the invention relating to the knowledge 4, 5, 6 and the knowledge 4, 5, 6 that are the basis of the present invention will be described.

First, as Knowledge 4, an example in which a living cell is placed on a tray and imaged using a multiphoton laser microscope (Olympus FV1000MPE) will be described. As a living cell, a living tissue picked up from the inside of a human body was used.

36A, 36B and 36C are images showing the cell morphology at a position 50 μm deep from the inner wall surface (mucosal surface) 113. FIG.

As the staining agent for staining the cell group, both staining agents including curcumin and staining agent including acid red (red No. 106) were used. The staining time was 5 minutes. In imaging, living cells were imaged at a temperature (37 ° C.) substantially the same as the body temperature.

36A to 36C are images obtained by simultaneously capturing the same cell group and extracting different colors (wavelengths) by applying a filter. FIG. 36A is an image obtained by extracting a color region stained with a curcumin pigment. FIG. 36B is an image obtained by extracting a color region stained with an acid red dye. FIG. 36C is an image obtained by extracting a color region stained with both the curcumin dye and the acid red dye. FIGS. 36A to 36C are black and white, but are originally color images. Due to the difference in dyeing tendency due to the staining agent, the area stained with curcumin dye is green fluorescent color, and the area stained with acid red dye is light red. It is represented by a fluorescent color close to orange, and the color difference is more clearly represented.

36A to 36C, the region indicated by the arrow I is a region in which the nuclei 135 of the gland cells 131 are arranged in a line along the basement membrane 137 and become normal cells. On the other hand, in the region indicated by the arrow II, two nuclei 135 exist between the center (lumen) of the gland 130 and the basement membrane 137. The region indicated by arrow II is not a malignant tumor, but is a region where canceration is about to begin.

FIG. 37 is a diagram showing a group of cancer cells in the stomach stained with curcumin dye. As shown in FIG. 37, the cancer cell group has a form in which the gland 130 and the nucleus 135 of the gland cell 131 cannot be identified.

In this way, by imaging the stained cell group using a multiphoton laser microscope, the gland 130, the basement membrane 137, the gland cells 131, and the nucleus 135 at a depth of 10 μm or more and 1000 μm or less from the inner wall surface (mucosal surface) 113 are detected. The form can be clearly understood. Then, by grasping the arrangement of the nuclei 135 in the gland cells 131, the distance between the basement membrane 137 and the nuclei 135, the shape and size of the nuclei 135, etc., pathological diagnosis of whether canceration has progressed can be accurately performed. Can be done.

On the other hand, multi-photon laser microscopes are generally expensive, and a method that enables more convenient pathological diagnosis for patients is desired. Therefore, the inventors tried to grasp these cell forms with a confocal laser microscope. In living body staining by applying a living body pigment to the mucosal surface, an example of imaging the tissue of the inner wall surface (mucosal surface) itself using a confocal laser microscope has been reported, but a depth of 20 μm or more from the inner wall surface 113 is reported. No example has been found in the past that makes it possible to image a cell group at a depth position.

Next, as knowledge 5, an example in which a cell morphology inside a living body is imaged using a confocal laser microscope (Olympus FV1000) will be described. A mouse was used as the living body.

As a stain for staining a cell group, a stain containing curcumin was used. The staining time was 5 minutes, which is longer than before. The dyeing time is preferably 3 minutes or more and 20 minutes or less. This is because if the staining time is shorter than 3 minutes, the staining agent does not penetrate into the cell tissue. Also, if the staining time exceeds 20 minutes, all cells are stained, making it difficult to distinguish between cancer cell groups and normal cell groups.

FIG. 38 is an image obtained by imaging a cell group inside the living body when viewed from a direction perpendicular to the inner wall surface (mucosal surface) 113 using a confocal laser microscope. The depth of the imaging region P from the inner wall surface (mucosal surface) 113 is about 5 μm in the region indicated by the arrow III and about 10 μm in the region indicated by the arrow IV. In the region indicated by the arrow III, the cytoplasm 126 of the epithelial cell 121 is dyed with a curcumin dye (green in an actual color image). In the region indicated by arrow IV, the cytoplasm 126 is dyed with curcumin pigment, and the nucleus 125 of the epithelial cell 121 is represented in black. Thus, since the shape of the nucleus 125 is almost the same size as the nucleus 125 of other regions and is not deformed, it can be seen that the region indicated by the arrow IV is a normal cell.

FIG. 39 is an image when a cell group inside the living body is imaged from the upper right side of the inner wall surface (mucosal surface) 113 using a confocal laser microscope. The depth of the imaging region P is about 5 μm in the region indicated by the arrow V, about 10 μm in the region indicated by the arrow VI, and about 50 μm in the region indicated by the arrow VII. In the region indicated by the arrow V, the cytoplasm 126 of the epithelial cell 121 is dyed with a curcumin dye (green in an actual color image). In the region indicated by arrow VI, the cytoplasm 126 is dyed with curcumin pigment, and the nucleus 125 of the epithelial cell 121 is represented in black. Since the shape of the nucleus 125 is almost the same size as the nucleus 125 in other regions and is not deformed, it can be seen that the region indicated by the arrow VI is a normal cell. In the region indicated by the arrow VII, the gland cells 131 are dyed with curcumin dye so that the gland cells 131 can be visually recognized.

FIG. 40 is an image when a cell group inside a living body is imaged from the upper left side with respect to the inner wall surface (mucosal surface) 113 using a confocal laser microscope. The depth of the imaging region P is about 5 μm in the region indicated by the arrow VIII and about 30 μm in the region indicated by the arrow IX. In the region indicated by the arrow IX, the cytoplasm 136 of the gland cell 131 is dyed with curcumin dye (green in the actual color image), and the outer periphery of the gland 130 and the position of the basement membrane 137 can be visually recognized. Further, the nucleus 135 of the gland cell 131 is shown in black. The plurality of nuclei 135 are arranged along the outer periphery while maintaining a substantially constant distance from the outer periphery of the basement membrane 137. Thus, since the nuclei 135 are regularly arranged in the gland 130, it can be seen that the region indicated by the arrow IX is a normal cell.

FIG. 41 is an image when a cell group inside a living body is imaged from the upper right side with respect to the inner wall surface (mucosal surface) 113 using a confocal laser microscope. The depth of the imaging region P is about 5 μm in the region indicated by the arrow X, about 30 μm in the region indicated by the arrow XI, and about 60 μm in the region indicated by the arrow XII. In the region indicated by the arrow XI, the cytoplasm 126 of the gland cells 131 is dyed with curcumin dye (green in the actual color image), and the outer periphery of the gland 130 and the position of the basement membrane 137 can be visually recognized. Arrows nuclei 135 are arranged along the outer periphery while maintaining a substantially constant distance from the outer periphery of the basement membrane 137. Thus, since the nuclei 135 are regularly arranged in the gland 130, it can be seen that the region indicated by the arrow XI is a normal cell. In the region indicated by the arrow XII, the capillary 132 is darkly dyed with curcumin dye.

Thus, even in the confocal laser microscope, the size of the

nuclei

125 and 135, the arrangement state of the

nuclei

125 and 135, the nuclei 135 and the basement membrane 137 can be adjusted by appropriately adjusting the focal position and the staining time. The suspicion of the lesion can be discriminated depending on whether the distance between the two is uniform.

Further, as Knowledge 6, another example in which the cell morphology inside the living body is imaged using a confocal laser microscope (Olympus FV1000) will be described. A mouse was used as the living body. The staining time of living cells with the staining agent was 5 minutes.

Furthermore, as a staining agent for staining a cell group, a staining agent containing curcumin with an optimized lysis method was used.

First, even if it can be dyed, uneven dyeing occurs, and it is difficult to obtain a homogeneous living body dyed image. Then, when the solvent which replaces water was examined, it turned out that curcumin is easy to melt | dissolve in ethanol which is tertiary alcohol and glycerol which is tertiary alcohol. In particular, curcumin is dissolved in 100% glycerol or 50% glycerol / 50% ethanol mixed solution in an amount of about 5%, so this was diluted and used for vital staining. Specifically, a 5% solution was used as a stock solution, and immediately before actual use, the stock solution was stained with a liquid diluted 10 to 1000 times with physiological saline. By optimizing this lysis method, a high-definition cell image as shown below could be obtained.

FIG. 42 is an image obtained by imaging a living cell stained with a stain optimized in lysis method with a confocal laser microscope, (a) is a normal colonic mucosa, (b) is an image showing colon cancer. It is. The depth of the imaging region is about 50 μm from the mucosal surface.

In the normal large intestine mucosa of FIG. 42 (a), the shape, size, and arrangement of the nuclei 135 are almost uniform, and the distance between the nuclei 135 and the basement membrane 137 is almost constant as indicated by the arrow XIII. Yes. Here, the nucleus is represented by a dark portion of sesame grains. Further, as in the region indicated by the arrow XIV, the distribution pattern of the structure (crypts) of the gland 130 is substantially uniform. Here, the crypt is represented by a dark portion near the center of the dashed circle. Further, the capillaries 132 show a regular running pattern around the crypts.

On the other hand, in the colorectal cancer of FIG. 42 (b), the shape, size and arrangement of glandular nuclei are non-uniform, and the distance between the glandular nuclei and the basement membrane is non-uniform, as in the region indicated by arrow XV. It has become. In addition, the structure of the gland (crypts) is not observed, and the capillaries have no regular crypts because they have no crypts. By analyzing the regularity of such nuclei and crypts on the image, the pathological diagnosis accuracy and diagnosis speed of cancer can be significantly improved. For example, the disorder of regularity can be detected by obtaining the approximate center of the nucleus or crypt from the image, connecting the centers with line segments, and comparing the lengths of the line segments. Further, even if the area of the regions connected by the line segments is obtained, the disorder of regularity can be similarly detected. For a short period of time, it can be useful for doctors to make a final diagnosis by categorizing detected irregularities in a group or creating a distribution map so that a group outside of a certain range is suspected of being cancerous. To be ready.

In this way, by optimizing the lysis method with biological staining using curcumin, it increases the uptake into cells and tissue penetration, stains the cells evenly, and using a confocal laser microscope, The cell structure could be clearly visualized.

To find the center from an image of the above-mentioned nucleus or crypt with a certain area, find the major axis and minor axis on the bitmap, then connect the major axis and minor axis with line segments and treat the intersection as the center. Can do. A crypt having a shape close to a line segment can be obtained on a bitmap with the midpoint of the line segment as the center. It is also possible to calculate by reducing the area of the image treated as a nucleus by utilizing the fact that the nucleus is regularly arranged around the crypt. The length of the line segment connecting these adjacent centers can be obtained on the bitmap. Although the figure is black and white, an actual image can obtain a dark part using color information by using fluorescence of staining. Furthermore, since the gland shown in FIG. 10A can be handled as a substantially circular image in normal cells, the disorder of regularity can be obtained digitally by utilizing the disorder of the regularity of the line segment connecting the centers of the glands. This calculation of irregularity of the regularity results in processing a huge amount of data, but the irregularity of regularity can be detected in a short time by using a computer.

Hereinafter, embodiments based on the findings 4, 5 and 6 will be described.

(Embodiment 4)
Next, the basic configuration of the laser endoscope apparatus 1C according to the fourth embodiment will be described with reference to FIGS. 43, 44, and 45. FIG.

FIG. 43 is a schematic diagram showing an end portion on the distal end side of the endoscope 2 of the laser endoscope apparatus 1C in FIG. FIG. 44 is a schematic diagram showing the entire endoscope 2. FIG. 45 is a block diagram showing a control configuration of the laser endoscope apparatus 1C.

As shown in FIG. 45, the laser endoscope apparatus 1 </ b> C includes an imaging unit 10 having an endoscope 2, a control unit 50, and an image processing unit 70. The laser endoscope apparatus 1 </ b> C includes a laser oscillator 60 and an optical component 65.

Further, the laser endoscope apparatus 1C includes a staining agent supply unit 40 that supplies the staining agent to the inside of the living body (see FIG. 12). In the present embodiment, the stain shown in Knowledge 5 or Knowledge 6 is used.

The laser L1 oscillated from the laser oscillator 60C is reflected by the dichroic mirror 66C, which is the optical component 65C, and further reflected by the mirror 19C in the endoscope 2 to irradiate the living body. The living cells irradiated with the laser L1 generate fluorescence, and the light due to the fluorescence is reflected by the mirror 19C, passes through the dichroic mirror 66C, and is detected by the photodetector 35C. The light detected by the photodetector 35C is converted into an electrical signal, and an image is formed by the image processing unit 70. Since the fluorescence color varies depending on the staining agent, a plurality of photodetectors 35C can be provided, and an optical filter that separates the colors can be placed in front of the photodetectors 35C to separate them. These operations and the functions and roles of each component are almost the same as those shown in FIG. 15, but since the confocal laser device is different in principle from the multiphoton laser device, “C” is added to each component number. Distinguish.

The laser oscillator 60C includes a plurality of types of lasers that can be stepwise varied in the wavelength range of 405 to 980 nm, and the wavelength is selected according to the characteristics of the fluorescence reaction to be measured. It may be pulse drive or continuous oscillation drive. In the case of pulse driving, a range in which a clear image can be obtained is selected in relation to the sweep frequency of imaging with a duty of 5% to 50% or more and several tens of kHz. The laser L1 in the present embodiment is a confocal laser, and the laser oscillator 60C uses, for example, a laser having a wavelength of 405 nm and an output of up to 30 mW. The laser output during imaging of this laser is emitted in the range of 5 to 10 mW, but is not limited to this. In the laser oscillator 60C, the intensity of the laser L1 can be adjusted according to the degree of staining and the degree of fluorescence.

The dichroic mirror 66C, which is the optical component 65, reflects the same wavelength as the laser L1 and transmits light of other wavelengths. Therefore, the laser L1 oscillated from the laser oscillator 60C is reflected toward the mirror 19C by the dichroic mirror 66C. On the other hand, the fluorescence generated in the living cells is reflected by the mirror 19C, passes through the dichroic mirror 66C, and reaches the photodetector 35C. The optical component 65C can also be configured by a prism, a 4 / λ plate, or the like.

The imaging unit 10 includes the endoscope 2 and the photodetector 35C, and images the cell morphology inside the living body by applying the laser L1 to the inside of the living body.

The photodetector 35C detects the fluorescence generated by applying the laser L1, and converts the fluorescence into an electrical signal corresponding to the fluorescence intensity. As the photodetector 35C, for example, a photomultiplier tube, a CCD semiconductor image sensor, or the like can be used. A pinhole is provided as a confocal laser function.

44. As shown in FIG. 44, the endoscope 2 includes an inner cylinder 12 and an outer cylinder 13 that surrounds a part of the inner cylinder 12. A part of the inner cylinder 12 and the outer cylinder 13 are inserted into the living body. The length of the inner cylinder 12 is, for example, 50 mm, and the outer diameter of the inner cylinder 12 is, for example, 3 to 10 mm. A linear actuator is attached to the inner cylinder 12, and the inner cylinder 12 can move about 25 mm in the axial direction X with respect to the outer cylinder 13. The inner cylinder 12 is equipped with an ultrasonic motor, and the inner cylinder 12 can rotate 360 ° with respect to the outer cylinder 13. The operation of the inner cylinder 12 in the axial direction X or the operation in the rotation direction R is controlled by the control unit 50.

An imaging head 11 is provided at the end of the endoscope 2 on the distal end side of the inner cylinder 12. As shown in FIG. 43, the imaging head 11 passes through the insertion tube 20 and is inserted into the living body together with the inner cylinder 12. The imaging head 11 is controlled to move inside the living body by operations in the axial direction X and the rotation direction R of the inner cylinder 12.

The imaging head 11 has an objective lens 16C, a focal point changing unit 18, a spacer 17, and a mirror 19C.

As described above, the mirror 19C changes the direction of the laser L1 output from the laser oscillator 60C toward the objective lens 16C, or changes the direction of the light fluorescent by the living cells toward the photodetector 35C. It is.

The objective lens 16C is provided to face the inner wall surface 113 of the living body. The objective lens 16 has, for example, a diameter of 10 mm, a magnification of 10 times, a resolution of 5 μm, and an imaging field of view of 3 mm × 3 mm. Alternatively, the objective lens 16 has a diameter of 12 mm, a magnification of 40 times, a resolution of 10 μm, and a visual field of 7.5 mm × 7.5 mm. The wider the field of view, the better. As the objective lens 16C, a lens having a diameter of 3 mm to 5 mm that can be easily inserted into a living body can be used as an objective lens that cuts a part of the lens having the diameter described on the left or obtains the same resolution.

It should be noted that the objective lens 16C may be arranged to be inclined with respect to the inner wall surface 113. By capturing an image with the objective lens 16C tilted, it is possible to simultaneously observe the cell morphology of both the epithelium 120 and the gland 130.

The focus variable unit 18 is, for example, a piezoelectric actuator or an electromagnetic actuator, and changes the focus position of the objective lens 16C by moving the objective lens 16C in the direction of the optical axis. The focus variable section 18 is controlled in operation by the control section 50 so that the focus can be adjusted from the inner wall surface (mucosal surface) 113 within a depth range of 0 to 75 μm. By changing the focal position, it is possible to image the state of the living body at a predetermined depth from the inner wall surface 113 of the digestive tract 112.

The spacer 17 is, for example, annular, and is provided around the space between the objective lens 16C and the inner wall surface 113. The spacer 17 is a component for preventing the objective lens 16C from touching the inner wall of the living body and for maintaining a constant distance between the objective lens 16C and the inner wall surface 113. The distance between the objective lens 16C and the inner wall surface (mucosal surface) 113 can be appropriately set within a range of, for example, 1 mm or more and 10 mm or less by replacing the spacer 17 before starting imaging or adding a mechanism that can be changed by an actuator or the like. Set to a value. The controller 50 controls the movement of the imaging head 11 (inner cylinder 12) while bringing the spacer 17 into contact with the inner wall surface 113, and maintains the distance of the objective lens 16C relative to the inner wall surface 113 constant.

The control unit 50 includes a CPU, a ROM, a RAM, and the like. The control unit 50 controls the operation of the imaging head 11 via the inner cylinder 12. Specifically, the control unit 50 controls the imaging head 11 to move in the circumferential direction along the inner circumference of the inner wall of the digestive tract 112, and in the duct direction (digestive tract axis) of the digestive tract 112. Move control along. Further, the control unit 50 controls the operation of the focus changing unit 18 to change the position of the objective lens 16C in the optical axis direction and control the focus position connected to the inside of the living body. The controller 50 can also adjust the laser output by controlling the laser oscillator 60C.

The image processing unit 70 stores the electrical signal (fluorescence intensity) converted by the photodetector 35C and the coordinate position of the imaging unit 10 sent from the control unit 50 in association with each other, and processes these data to perform digital processing. Generate an image. The generated digital image is displayed on a monitor, printed out, or recorded in a storage device, for example. As an example of the coordinate position of the imaging unit 10, a distance from a location (for example, throat or anus) serving as a reference for the patient, a rotation angle of the imaging head 11, and the like can be used.

The confocal laser endoscope apparatus 1C according to the present embodiment includes an imaging head 11 that is inserted into a living body, and captures an image of the living body by applying a laser to the living body via the imaging head 11. And a control unit 50 that controls the operation of the imaging head 11. The imaging head 10 includes an objective lens 16C, and a focus variable unit 18 that can change the focal position of the objective lens 16C in the depth direction of the living body, and the control unit 50 has a focal position in the mucous membrane inside the living body. The focus varying unit 18 is operated so as to have a predetermined depth within a depth of 10 μm or more and 100 μm or less (preferably 10 μm or more and 70 μm or less) from the surface, and the imaging unit 10 selectively has a cell group inside the living body. A laser is applied to the stained cell group in contact with the staining agent to be stained for at least 2 minutes, preferably 5 minutes or more, and the stained cell group at a predetermined depth is imaged. Here, a method for controlling the focus while keeping the positions of the objective lens 16C and the mucosal surface constant will be described. Reference numeral 171 in FIG. 45 denotes a second laser oscillator that oscillates continuous parallel light as reference light having a wavelength of about 680 nm and an output of about 5 mW, for example. A beam splitter, a half mirror, or the like is inserted into the same optical path as the laser oscillator 60C. In FIG. 45, for easy understanding, the optical path L2 is indicated by a broken line slightly shifted in position. The reference light L 2 follows substantially the same path as the inspection laser light L 1, but the optical path is changed by the beam splitter 173 and enters the focus control optical unit 174. Here, the optical lens configuration is such that when the focal position of the objective lens 16C is changed by a cylindrical lens and a beam splitter, the amount of change can be detected. 175 is a photodetector, and the light detected by the photodetector usually divided into 2 or 4 blocks is converted into an electrical signal proportional to the relative position fluctuation of the objective lens 16C and the mucous membrane surface by a differential amplifier or the like. . Such position control of the objective lens is used in an optical disc apparatus or the like, and can be sufficiently applied to an endoscope apparatus. Here, it should be noted that the endoscope apparatus preferably has different wavelengths so that the imaging laser beam L1 and the reference beam L2 can be easily separated. By separating the wavelength by 100 nm or more, it is possible to obtain the optical characteristics of the imaging system and the focus control system with good separation characteristics. Further, when the focus control system as described above is provided, the focus position can be finely adjusted by applying a bias voltage in the control system. By changing this bias voltage stepwise, the focal position of the laser beam L1 can be automatically controlled in the depth direction.

The

optical parts

11C, 35C, 65C, 66C, 172, 173, and 174 are greatly affected by the transmittance and reflectance depending on the L1 and L2 laser wavelengths. By preparing, even when the laser wavelength is changed depending on the staining agent to be used or the site to be inspected, it can be easily handled.

Thus, even in the confocal laser endoscope apparatus 1C, an image at a depth of 10 μm or more and 70 μm or less can be acquired from the inner wall surface (mucosal surface) 113 of the living body by taking sufficient staining time. It is possible. Thereby, a lesion can be easily found, and an image can be acquired without applying a laser load to the patient by selecting a wavelength and a laser intensity.

(Other examples)
Although the laser endoscope apparatuses 1 to 1C according to the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments and modifications thereof. For example, aspects in which the following modifications are made to the above-described embodiment and its modifications can also be included in the present invention.

In the above-described embodiment, the case of staining with a staining agent such as curcumin has been described. On the other hand, it is possible to visualize the morphology of cells within a depth of 10 μm to 1000 μm from the mucosal surface and to detect cancer without missing the entire pancreatic image of the gastrointestinal tract. Is also possible. Here, the cell morphology includes the cytoplasm of each cell, the shape of the nucleus, the arrangement pattern of the crypts of the gland, the capillary running pattern, and the like. The above-described detection is possible because a certain amount of fluorescence is emitted by laser excitation of intracellular chemical substances FAD (flavin adenine dinucleotide), NAD (nicotinamide adenine dinucleotide) and the like. This is the same whether observed with a confocal laser microscope or a multiphoton laser microscope. However, the problem is that it is necessary to irradiate a large amount of excitation laser light. It is about 20 times or more the amount of light necessary for imaging after living body staining using curcumin etc., and there is a possibility that damage to living cells may increase, but it can be improved by increasing the sensitivity of the detection system It is.

As another embodiment, FIG. 46 shows a case of observation with a confocal laser microscope.

FIG. 46 is a photograph taken on the inner surface of an unstained mouse large intestine mucosa using a confocal laser microscope. First, an image (a) of a surface at an arbitrary location and an image (b) inside 10 μm from the surface of the location are shown in the upper and lower left columns. Also, an image (c) of the surface at a location displaced by about 100 μm from the above location and an image (d) inside 10 μm from the surface of the location are shown. Further, an image (d) obtained by combining and joining the image (b) and the image (d) inside 10 μm using the crypt arrangement pattern as a mark is shown. Here, the scale bar is 100 μm.

Next, FIG. 47 shows the case of observation with a multiphoton laser microscope.

FIG. 47 is an image taken using a multiphoton laser microscope on the unstained mouse large intestine mucosa inner surface. First, the image (a) of the surface at an arbitrary place and the image (b) inside 25 μm from the surface of the place are shown in the left column. The right column shows an image (c) of the surface at a position shifted by about 400 μm from the above-mentioned place and an image (d) inside 25 μm from the surface of the place. Further, an image (e) obtained by joining the image (b) and the image (d) inside 25 μm using the arrangement pattern of the crypts as a mark is shown. By performing this continuously, panorama can be realized. An arrow e1 in the image (b) and a star e2 in the image (d) correspond to the arrow e1 and the star e2 in the synthesized image (e). The scale bar in the figure is 100 μm. In addition, in the image (f) obtained by photographing the image (e) at a zoom magnification of 2 times, the cytoplasm of epithelial cells and glandular cells appears bright. Further, the portion of the nucleus 135 indicated by the arrow looks dark. The scale bar of the image (f) is 100 μm.

As shown here, even in the case of no staining, if the nucleus and crypts can be grasped image-wise, the cancer detection method shown in the embodiment in which cells are stained with curcumin or the like can be used. is there. It may be a comparison of cell shape and brightness, or a line segment using a nucleus or crypt on the image or a comparison of areas surrounded by the line segment.

For example, in the present embodiment, when performing double dyeing, dyeing is performed sequentially using one type of dyeing agent. However, the present invention is not limited thereto, and a plurality of dyes are mixed in advance to create a mixed dyeing agent containing both, You may dye | stain simultaneously using a mixed dyeing agent.

In the present embodiment, the living body is stained and imaged by double staining using two color stains, but the present invention is not limited thereto, and the living body is stained and imaged by multiple staining using two or more colorants. It is also possible.

For example, when performing multiple staining of three or more colors on a cell group of a living body, the following method may be mentioned. First, dyeing agent A is collected after dyeing with dyeing agent A containing dye A1. Then, after dyeing | staining using the dye B containing the pigment | dye B1, the dye B is collect | recovered. Then, after dyeing | staining using the dyeing agent C1 containing the pigment | dye C1, the dyeing | staining agent C1 is collect | recovered. Thereby, multiple staining can be performed. In this case, since each color surely touches the cell group, the staining property of each cell can be improved.

In addition, as another method for performing multiple staining, there is a method in which a mixed solution ABC in which a plurality of dyes A1, B1, and C1 are mixed in advance is prepared, and the mixed solution ABC is used for staining. According to this, dyeing in a short time becomes possible.

Furthermore, it is also within the scope of the present invention to simply stain the oral cleaning liquid to be treated by including a mucosal cleaning agent or a staining agent before the inspection using the laser endoscope apparatus.

For example, in the present embodiment, a multiphoton laser is used as the laser of the laser endoscope apparatus 1, but the present invention is not limited to this, and a confocal laser 1C can also be used.

In addition, the spacer 17 of the imaging head 11 in the present embodiment is not limited to an annular shape, and may be a plurality of members provided so as to surround a space between the objective lens 16 and the inner wall surface 113, or an objective. A pair of members that sandwich a space between the lens 16 and the inner wall surface 113 may be used.

In addition, when the living body moves when imaging the inside of the living body, the focus variable unit 18 is focus-controlled using the above-described reference laser light or the like using the control unit 50, and the image is captured while being focused. Also good. Further, the control unit 50 may process the wobble signal for driving the objective lens in a sine wave shape or a staircase wave shape at a constant period and the image to process the image while matching the imaging position.

In the present embodiment, imaging is performed while specifying the depth position from the inner wall surface (mucosal surface) 113 of the digestive tract 112, the depth information and the image information are stored in correspondence with each other, and the same depth position is stored. The synthesized image is generated by synthesizing the captured images. However, the present invention is not limited to this. For example, without recognizing the depth position, a plurality of images having different depth positions and imaging regions P are acquired, and a similar image or a connected image is extracted from the plurality of images and combined to generate a composite image May be.

In the present embodiment, cells inside the living body are stained with a staining agent and then imaged using the laser endoscope apparatus 1. However, the present invention is not limited to this, and a multiphoton laser can be used without staining with a staining agent. By using the laser endoscope apparatus according to, it is possible to take an image of the cell morphology inside the living body. For example, when irradiated with a multiphoton laser, light having a wavelength half that of the multiphoton laser is generated in the cell by a compound (such as NAD: nicotinamide adenine dinucleotide) that exists universally in the cell, and the generated light is NAD. Since it is self-fluorescent upon hitting a compound such as, an image of the cell morphology inside the living body can be obtained without exogenous staining.

Further, the duct direction (axial direction) of the digestive tract is not limited to a straight line, and the present invention can be applied to a curved line.

In addition, the laser endoscope apparatuses 1 to 1C in the present embodiment can be applied to luminal organs (bronchi, bladder, ureter, etc.) other than the digestive tract, and within 1 mm from the surface. Although there are limitations, cell structures such as kidney, liver, brain, and retina can be visualized.

In the above-described embodiment, the laser microscope and the laser endoscope are described. However, the microscope function is used for imaging and diagnosis of the epidermis, and the microscope function is used for imaging and diagnosis of the internal organs such as the digestive tract. It is handled as an endoscope having.

The laser endoscope apparatus according to the present invention is used for imaging and treating a wide range of lesions occurring in the digestive tract, respiratory tract, renal urinary tract, uterine ovarian genitalia, and cerebrospinal nerve without leakage. .

DESCRIPTION OF

SYMBOLS

1, 1A, 1B, 1C Laser endoscope apparatus 2 Endoscope 10 Imaging part 11 Imaging head 12 Inner cylinder 13 Outer cylinder 15

Arm

16, 16C Objective lens 17 Spacer 17a Wheel 18 Focus

variable part

19, 19C Mirror 20 Insertion tube 21 first balloon 22 second balloon 23 pressing member 24 support roller 25 expansion mechanism 26 sliding

member

27, 28 joint mechanism 29 gyro sensor 30 pressure sensor 31

telescopic spacer

35, 35C photodetector 40 staining agent supply unit 42 supply port 42a Discharge port 43 Collection port 45 Stain 50 Control unit 51 First focus variable mode 52 Second

focus variable mode

60, 60C Laser oscillator 65,

65C Optical component

66, 66C Dichroic mirror 70 Image processing unit 81 Angle detector 82 Linear Scale 102 Multiphoton laser microscope 11 The inner wall surface of the gastrointestinal tract 113 digestive tract (mucosal surface)
113a Uneven part 114 Gastrointestinal axis 120 Epithelium 121 Epithelial cell 125 Epithelial cell nucleus 126 Epithelial cell cytoplasm 130 Gland 131 Gland cell 132 Capillary 133 Connective tissue 135 Gland cell nucleus 136 Gland cell cytoplasm 137 Basement membrane 138 Crypt
152 Cancer cell population 160 Mucosal fascia A, B Staining agents P, P1, P2, P3 Imaging regions Pa, Pb Regions where imaging regions overlap L Laser R Circumferential direction (rotation direction)
S Closed space X Axial direction

Claims

An imaging unit having an imaging head inserted into a living body, and imaging the living body by applying a laser to the living body via the imaging head;
A control unit for controlling the imaging head to move inside the living body;
An image processing unit that processes an image captured by the imaging unit,
The imaging unit images a plurality of imaging areas to be imaged with movement of the imaging head so that a part of adjacent imaging areas overlaps,
The image processing unit generates a composite image by superimposing regions where the plurality of imaging regions overlap each other.
The imaging unit images the imaging region at a predetermined depth out of a depth of 10 μm or more and 1000 μm or less from the mucosal surface inside the living body,
The laser endoscope apparatus according to claim 1, wherein the image processing unit generates the composite image at the predetermined depth.
The laser endoscope apparatus according to claim 1, wherein the control unit controls the movement of the imaging head so that the imaging head scans in a state where the imaging head maintains a certain distance from the living body.
The imaging head includes an objective lens disposed to face the living body, and a spacer provided around a space between the objective lens and the living body,
The laser endoscope apparatus according to claim 3, wherein the control unit maintains the certain distance by controlling the movement of the imaging head so that the spacer contacts the living body.
The living body is a digestive tract;
The control unit controls the imaging head to move along the inner periphery of the digestive tract,
The imaging unit images a plurality of imaging areas to be imaged with the movement of the imaging head so that a part of imaging areas adjacent in the circumferential direction overlaps,
The laser endoscope apparatus according to any one of claims 1 to 4, wherein the image processing unit generates a panoramic image by superimposing regions where the plurality of imaging regions overlap each other.
The living body is a digestive tract;
The control unit controls the imaging head to rotate about the axis of the digestive tract;
The imaging unit images a plurality of imaging areas to be imaged with rotation of the imaging head so that a part of imaging areas adjacent to each other in the rotation direction overlaps,
The laser endoscope apparatus according to any one of claims 1 to 4, wherein the image processing unit generates a panoramic image by superimposing regions where the plurality of imaging regions overlap each other.
The laser endoscope apparatus according to claim 6, wherein the control unit controls the imaging head to revolve around an axis of the digestive tract.
The laser endoscope apparatus according to claim 7, wherein the control unit controls the imaging head to move in a spiral direction around an axis of the digestive tract.
The control unit controls the imaging head so as to move along a duct direction of the digestive tract,
The imaging unit images a plurality of imaging regions to be imaged with the movement of the imaging head so that a part of the imaging regions adjacent in the pipe line direction overlaps,
The laser endoscope apparatus according to any one of claims 5 to 8, wherein the image processing unit generates the panoramic image by superimposing regions where the plurality of imaging regions overlap each other.
The imaging head includes an objective lens, and a focus variable unit capable of changing a focal position of the objective lens in a depth direction from a cell surface of the living body,
The control unit changes the focus position by operating the focus variable unit,
The imaging unit images a plurality of imaging regions having different depths with the change of the focal position,
The image processing unit obtains a stereoscopic image inside the living body by arranging a plurality of images obtained by imaging of the imaging unit in correspondence with the focal position. The laser endoscope apparatus described in 1.
The control unit is configured to change the focus position at a first pitch, a first variable focus mode, and a second focus at which the focus position is changed at a second pitch that is smaller than the first pitch. A portion suspected of having a lesion when a portion suspected of having a lesion exists in the image obtained by the imaging after performing the imaging in the first variable focus mode. The laser endoscope apparatus according to claim 10, wherein the imaging is performed in the second variable focus mode in the vicinity of a focal position when the image of the above is captured.
The control unit stores in advance an image of normal cells in the absence of a lesion, and the image obtained in the first variable focus mode and the image of the normal cells are obtained for at least one of shape and brightness. The laser endoscope apparatus according to claim 11, wherein the suspicion of the lesion is determined by comparison.
The control unit, when there is a diseased cell in the image obtained by the imaging unit, raise the output of the laser than at the time of imaging, hit the laser that raised the output to the diseased cell, The laser endoscope apparatus according to any one of claims 1 to 12, wherein the diseased cells are removed.
The laser endoscope apparatus according to any one of claims 1 to 13, wherein the laser is a multiphoton laser.
further,
A staining agent supply unit for supplying a staining agent for selectively staining a cell group inside the living body to a chromatic color, and supplying the inside of the living body;
The laser endoscope apparatus according to any one of claims 1 to 14, wherein the imaging unit images the cell group stained by the staining agent supply unit.
further,
A staining agent supply unit for supplying a staining agent for staining a cell group inside the living body into two or more selective chromatic colors depending on the type of cells, and supplying the inside of the living body;
The laser endoscope apparatus according to any one of claims 1 to 14, wherein the imaging unit images the cell group stained with two or more colors by the staining agent supply unit.
A staining agent supply unit for supplying a staining agent for staining a cell group inside the living body to two or more selective chromatic colors depending on the type of the cell;
A laser endoscope apparatus comprising: an imaging unit that images the cell group by applying a laser to the cell group stained by the staining agent supply unit.
The staining agent includes two staining agents including a staining agent containing both curcumin (AccumRed) and a staining agent including Curcumin, and a staining agent including Acid Red. The laser endoscope apparatus according to claim 17.
The staining agent includes two types of staining agents including both curcumin (Curcumin) and Fast Green FCF (FastGreen FCF), or a staining agent including Curcumin (Curcumin) and a staining agent including Fast Green FCF (FastGreen FCF). The laser endoscope apparatus according to claim 17, which is a staining agent.
Supplying the inside of the living body with a staining agent for specifically staining a chromatic color around the cancer cell surrounding cells other than the cancer cells located in the periphery of the cancer cell among the inside cells of the living body A staining agent supply unit,
A laser endoscope apparatus comprising: an imaging unit that captures an image that allows the cancer cell peripheral cell group to be visually discriminated by applying a laser to the cell group inside the living body stained by the stain supply unit.
The laser endoscope apparatus according to claim 20, wherein the stain is a stain containing Rose Bengal.
An imaging unit having an imaging head inserted into a living body, and imaging the living body by applying a laser to the living body via the imaging head;
A control unit for controlling the operation of the imaging head,
The imaging head includes an objective lens, and a focus variable unit capable of changing a focal position of the objective lens in a depth direction from a cell surface of the living body,
The control unit changes the focus position by operating the focus variable unit,
The imaging unit images a plurality of imaging regions having different depths from a mucosal surface inside the living body in accordance with the change of the focal position.
The imaging unit images the imaging region in a predetermined range of depths from 0 μm to 1000 μm from the mucosal surface inside the living body, and stores the captured image and the depth information in association with each other. ,
An image processing unit for processing an image captured by the imaging unit;
The laser endoscope according to claim 22, wherein the image processing unit generates a stereoscopic image inside the living body by arranging a plurality of images obtained by imaging of the imaging unit in correspondence with the focal position. Mirror device.
further,
A staining agent supply unit for supplying a staining agent for selectively staining a cell group inside the living body to a chromatic color, and supplying the inside of the living body;
The laser endoscope apparatus according to claim 23, wherein the imaging unit images the cell group stained by the staining agent supply unit.
further,
A staining agent supply unit for supplying a staining agent for staining a cell group inside the living body into two or more selective chromatic colors depending on the type of cells, and supplying the inside of the living body;
The laser endoscope apparatus according to claim 23, wherein the imaging unit images the cell group stained with two or more colors by the staining agent supply unit.
further,
Supplying the inside of the living body with a staining agent for specifically staining a chromatic color around the cancer cell surrounding cells other than the cancer cells located in the periphery of the cancer cell among the inside cells of the living body A staining agent supply unit
The laser endoscope apparatus according to claim 23, wherein the imaging unit images the cancer cell peripheral cell group stained by the staining agent supply unit.
The image processing unit generates a cross-sectional image of the stained cell group by cutting the plurality of images at a position including the stained cell group,
The laser endoscope according to any one of claims 24 to 26, wherein the control unit determines a suspicious lesion based on a depth at which the cell group represented in the cross-sectional image is stained. apparatus.